U.S. patent application number 17/630138 was filed with the patent office on 2022-09-08 for discovery, total synthesis, and bioactivity of doscadenamides.
This patent application is currently assigned to University of Florida Research Foundation, Incorporation. The applicant listed for this patent is Smithsonian Institution, University of Florida Research Foundation, Incorporation. Invention is credited to Qi-Yin Chen, Jason C. Kwan, Xiao Liang, Hendrik Luesch, Susan Matthew, Valerie J. Paul.
Application Number | 20220281816 17/630138 |
Document ID | / |
Family ID | 1000006392211 |
Filed Date | 2022-09-08 |
United States Patent
Application |
20220281816 |
Kind Code |
A1 |
Luesch; Hendrik ; et
al. |
September 8, 2022 |
DISCOVERY, TOTAL SYNTHESIS, AND BIOACTIVITY OF DOSCADENAMIDES
Abstract
The invention is directed towards compounds (e.g., Formulae
(I)-(IX)), their mechanism of action, processes to prepare the
compounds, methods of activating quorum sensing signaling activity,
and methods of treating diseases and disorders using the compounds
described herein (e.g., Formulae (I)-(IX)).
Inventors: |
Luesch; Hendrik;
(Gainesville, FL) ; Liang; Xiao; (Gainesville,
FL) ; Matthew; Susan; (Houston, TX) ; Kwan;
Jason C.; (Gainesville, FL) ; Chen; Qi-Yin;
(Gainesville, FL) ; Paul; Valerie J.; (Fort
Pierce, FL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Florida Research Foundation, Incorporation
Smithsonian Institution |
Gainesville
Washington |
FL
DC |
US
US |
|
|
Assignee: |
University of Florida Research
Foundation, Incorporation
Gainesville
FL
Smithsonian Institution
Washington
DC
|
Family ID: |
1000006392211 |
Appl. No.: |
17/630138 |
Filed: |
July 24, 2020 |
PCT Filed: |
July 24, 2020 |
PCT NO: |
PCT/US20/43495 |
371 Date: |
January 25, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62879246 |
Jul 26, 2019 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 207/38
20130101 |
International
Class: |
C07D 207/38 20060101
C07D207/38 |
Goverment Interests
GOVERNMENT SUPPORT INFORMATION
[0002] This invention was made with government support under Grant
Nos. CA172310 and GM086210 awarded by the National Institutes of
Health. The government has certain rights in the invention.
Claims
1. A compound of Formula (I): ##STR00189## or a pharmaceutically
acceptable salt thereof; wherein R.sub.1 is H, Boc, acetyl, Fmoc,
or ##STR00190## R.sub.2 is H or C.sub.1-C.sub.6 alkyl; R.sub.3 is
H; R.sub.4 is H or C.sub.1-C.sub.6 alkyl; R.sub.5 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; and R.sub.7 is
H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl.
2. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.1 is ##STR00191##
3. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein R.sub.2 is C.sub.1-C.sub.6 alkyl.
4. The compound of claim 3, or a pharmaceutically acceptable salt
thereof, wherein R.sub.2 is Me.
5. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkynyl.
6. The compound of claim 5, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00192##
7. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkenyl.
8. The compound of claim 7, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00193##
9. The compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.1-C.sub.6 alkyl.
10. The compound of claim 9, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00194##
11. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H.
12. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H and R.sub.5 is C.sub.1-C.sub.6
alkyl.
13. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H and R.sub.5 is Me.
14. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H.
15. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H and R.sub.4 is C.sub.1-C.sub.6
alkyl.
16. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H and R.sub.4 is Me.
17. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkynyl.
18. The compound of claim 17, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00195##
19. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkenyl.
20. The compound of claim 19, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00196##
21. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.1-C.sub.6 alkyl.
22. The compound of claim 21, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00197##
23. The compound of any one of claims 1-22, or a pharmaceutically
acceptable salt thereof, according to Formula (II): ##STR00198## or
a pharmaceutically acceptable salt thereof.
24. The compound of claim 23, or a pharmaceutically acceptable salt
thereof, according to Formula (III): ##STR00199## or a
pharmaceutically acceptable salt thereof.
25. The compound of claim 24, or a pharmaceutically acceptable salt
thereof, according to Formula (IV): ##STR00200## or a
pharmaceutically acceptable salt thereof.
26. The compound of any one of claims 1-25, or a pharmaceutically
acceptable salt thereof, wherein the compound is: ##STR00201## or a
pharmaceutically acceptable salt thereof.
27. A compound of Formula (V): ##STR00202## or a pharmaceutically
acceptable salt thereof; wherein R.sub.4 is H or C.sub.1-C.sub.6
alkyl; R.sub.5 is H or C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00203## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
28. The compound of claim 27, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is ##STR00204##
29. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H.
30. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is C.sub.1-C.sub.6 alkyl.
31. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6
alkyl.
32. The compound of claim 30 or 31, or a pharmaceutically
acceptable salt thereof, wherein R.sub.10 is Me.
33. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H.
34. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is C.sub.1-C.sub.6 alkyl.
35. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6
alkyl.
36. The compound of claim 34 or 35, or a pharmaceutically
acceptable salt thereof, wherein R.sub.9 is Me.
37. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkynyl.
38. The compound of claim 37, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00205##
39. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkenyl.
40. The compound of claim 39, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00206##
41. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.1-C.sub.6 alkyl.
42. The compound of claim 41, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00207##
43. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H.
44. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H and R.sub.5 is C.sub.1-C.sub.6
alkyl.
45. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.4 is H and R.sub.5 is Me.
46. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H.
47. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H and R.sub.4 is C.sub.1-C.sub.6
alkyl.
48. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.5 is H and R.sub.4 is Me.
49. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkynyl.
50. The compound of claim 49, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00208##
51. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkenyl.
52. The compound of claim 51, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00209##
53. The compound of claim 28, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.1-C.sub.6 alkyl.
54. The compound of claim 53, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00210##
55. The compound of any one of claims 28-54, or a pharmaceutically
acceptable salt thereof, wherein the compound is: ##STR00211## or a
pharmaceutically acceptable salt thereof.
56. A compound of Formula (VI): ##STR00212## or a pharmaceutically
acceptable salt thereof; wherein Ru is H, Boc, acetyl, Fmoc, or
##STR00213## R.sub.11 is C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00214## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
57. The compound of claim 56, or a pharmaceutically acceptable salt
thereof, wherein R.sub.11 is C.sub.1-C.sub.6 alkyl.
58. The compound of claim 57, or a pharmaceutically acceptable salt
thereof, wherein R.sub.11 is Me.
59. The compound of claim 56, or a pharmaceutically acceptable salt
thereof, wherein R.sub.8 is ##STR00215##
60. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H.
61. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is C.sub.1-C.sub.6 alkyl.
62. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6
alkyl.
63. The compound of claim 61 or 62, or a pharmaceutically
acceptable salt thereof, wherein R.sub.10 is Me.
64. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H.
65. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is C.sub.1-C.sub.6 alkyl.
66. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6
alkyl.
67. The compound of claim 65 or 66, or a pharmaceutically
acceptable salt thereof, wherein R.sub.9 is Me.
68. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkynyl.
69. The compound of claim 68, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00216##
70. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkenyl.
71. The compound of claim 70, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00217##
72. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.1-C.sub.6 alkyl.
73. The compound of claim 72, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00218##
74. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkynyl.
75. The compound of claim 74, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00219##
76. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkenyl.
77. The compound of claim 76, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00220##
78. The compound of claim 59, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is C.sub.1-C.sub.6 alkyl.
79. The compound of claim 78, or a pharmaceutically acceptable salt
thereof, wherein R.sub.6 is ##STR00221##
80. The compound of any one of claims 58-79, or a pharmaceutically
acceptable salt thereof, wherein the compound is: ##STR00222## or a
pharmaceutically acceptable salt thereof.
81. A compound of Formula (VII): ##STR00223## or a pharmaceutically
acceptable salt thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc,
##STR00224## R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H
or C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00225## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
82. The compound of claim 81, or a pharmaceutically acceptable salt
thereof, wherein R.sub.8 is ##STR00226##
83. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H.
84. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is C.sub.1-C.sub.6 alkyl.
85. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6
alkyl.
86. The compound of claim 84 or 85, or a pharmaceutically
acceptable salt thereof, wherein R.sub.10 is Me.
87. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H.
88. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.9 is C.sub.1-C.sub.6 alkyl.
89. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6
alkyl.
90. The compound of claim 88 or 89, or a pharmaceutically
acceptable salt thereof, wherein R.sub.9 is Me.
91. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkynyl.
92. The compound of claim 91, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00227##
93. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.2-C.sub.6 alkenyl.
94. The compound of claim 93, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00228##
95. The compound of claim 82, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is C.sub.1-C.sub.6 alkyl.
96. The compound of claim 95, or a pharmaceutically acceptable salt
thereof, wherein R.sub.7 is ##STR00229##
97. The compound of claim 81, or a pharmaceutically acceptable salt
thereof, wherein R.sub.13 is ##STR00230##
98. The compound of claim 87, or a pharmaceutically acceptable salt
thereof, wherein R.sub.14 is H.
99. The compound of claim 97, or a pharmaceutically acceptable salt
thereof, wherein R.sub.14 is H and R.sub.15 is C.sub.1-C.sub.6
alkyl.
100. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.14 is H and R.sub.15 is Me.
101. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.15 is H.
102. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.15 is H and R.sub.14 is C.sub.1-C.sub.6
alkyl.
103. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.15 is H and R.sub.10 is Me.
104. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkynyl.
105. The compound of claim 104, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is ##STR00231##
106. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is C.sub.2-C.sub.6 alkenyl.
107. The compound of claim 106, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is ##STR00232##
108. The compound of claim 97, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is C.sub.1-C.sub.6 alkyl.
109. The compound of claim 108, or a pharmaceutically acceptable
salt thereof, wherein R.sub.6 is ##STR00233##
110. The compound of any one of claims 81-109, or a
pharmaceutically acceptable salt thereof, wherein the compound is:
##STR00234## or a pharmaceutically acceptable salt thereof.
111. A compound of Formula (VIII): ##STR00235## or a
pharmaceutically acceptable salt thereof; wherein R.sub.16 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; and R.sub.17 is H or C.sub.1-C.sub.6 alkyl; or R.sub.16 is
H or C.sub.1-C.sub.6 alkyl; and R.sub.17 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl.
112. The compound of claim 111, or a pharmaceutically acceptable
salt thereof, wherein the compound is ##STR00236## (doscadenamide
S4), or a pharmaceutically acceptable salt thereof.
113. A pharmaceutical composition comprising a compound of any one
of claims 1-112, or a pharmaceutically acceptable salt thereof, and
a pharmaceutically acceptable carrier.
114. The composition of claim 113, further comprising an additional
therapeutic agent.
115. The composition of claim 114, wherein the additional
therapeutic agent is an antibacterial agent.
116. The composition of claim 114, wherein the additional
therapeutic agent is an anticancer agent.
117. The composition of claim 114, wherein the additional
therapeutic agent is tumor necrosis factor-related
apoptosis-inducing ligand (TRAIL) or a TRAIL receptor agonist.
118. A method of activating quorum sensing signaling comprising the
administration of a compound of Formula (IX): ##STR00237## or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, ##STR00238##
each R.sub.14 is independently H or C.sub.1-C.sub.6 alkyl; R.sub.15
is H or C.sub.1-C.sub.6 alkyl; each R.sub.6 is independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00239## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
119. A method of activating quorum sensing signaling comprising the
administration of a compound of Formula (VII): ##STR00240## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, ##STR00241## R.sub.14 is H or C.sub.1-C.sub.6
alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00242## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
120. The method of claim 118 or 119, wherein the compound is
administered in vitro.
121. The method of claim 118 or 119, wherein the compound is
administered in vivo.
122. A method of activating quorum sensing signaling in a subject,
the method comprising the administration to the subject a compound
of Formula (IX): ##STR00243## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, or ##STR00244##
R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00245## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
123. A method of activating quorum sensing signaling in a subject,
the method comprising the administration to the subject a compound
of Formula (IX): ##STR00246## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, or ##STR00247##
R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00248## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
124. A method of inhibiting bacterial growth comprising the
administration of a compound of Formula (IX): ##STR00249## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, or ##STR00250## R.sub.14 is H or C.sub.1-C.sub.6
alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00251## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
125. The method of claim 120, wherein the inhibition is in
vitro.
126. The method of claim 120, wherein the inhibition is in
vivo.
127. A method of inhibiting bacterial growth in a subject, the
method comprising the administration to the subject a compound of
Formula (IX): ##STR00252## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, or ##STR00253##
R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00254## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
128. A method of treating a bacterial infection in a subject, the
method comprising the administration to the subject a compound of
Formula (VII): ##STR00255## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, ##STR00256##
R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00257## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
129. The method of claim 127 or 128, wherein the bacterial
infection is mediated by one or more Gram negative bacteria.
130. The method of claim 127 or 128, wherein the bacterial
infection is mediated by one or more Gram positive bacteria.
131. The method of claim 127 or 128, wherein the bacterial
infection is mediated by Pseudomonas aeruginosa.
132. A method of treating cancer in a subject, the method
comprising administering an effective amount of a compound of
Formula (IX): ##STR00258## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, ##STR00259##
each R.sub.14 is independently H or C.sub.1-C.sub.6 alkyl; R.sub.15
is H or C.sub.1-C.sub.6 alkyl; each R.sub.6 is independently H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00260## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
133. A method of treating cancer in a subject, the method
comprising administering an effective amount of a compound of
Formula (VII): ##STR00261## or a pharmaceutically acceptable salt
thereof; wherein R.sub.13 is H, Boc, acetyl, Fmoc, ##STR00262##
R.sub.14 is H or C.sub.1-C.sub.6 alkyl; R.sub.15 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H,
Boc, acetyl, Fmoc, or ##STR00263## R.sub.7 is H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
134. A method of increasing the effectiveness of anti-cancer
therapy in a subject currently being administered one or more
anti-cancer therapies, the method comprising administering an
effective amount of a compound of Formula (IX): ##STR00264## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, ##STR00265## each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; each
R.sub.6 is independently H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H, Boc, acetyl,
Fmoc, or ##STR00266## R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9 is H
or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
135. A method of increasing the effectiveness of anti-cancer
therapy in a subject currently being administered one or more
anti-cancer therapies, the method comprising administering an
effective amount of a compound of Formula (VII): ##STR00267## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, ##STR00268## R.sub.14 is H or C.sub.1-C.sub.6
alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00269## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl.
136. The method of claim 134 or 135, wherein the anti-cancer
therapy is tumor necrosis factor-related apoptosis-inducing ligand
(TRAIL) or a TRAIL receptor agonist.
137. A method of increasing the effectiveness of anti-bacterial
therapy in a subject currently being administered one or more
anti-bacterial therapies, the method comprising administering an
effective amount of a compound of Formula (IX): ##STR00270## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, ##STR00271## each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; each
R.sub.6 is independently H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.8 is H, Boc, acetyl,
Fmoc, or ##STR00272## R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.9 is H
or C.sub.1-C.sub.6 alkyl; and R.sub.10 is H or C.sub.1-C.sub.6
alkyl.
138. A method of increasing the effectiveness of anti-bacterial
therapy in a subject currently being administered one or more
anti-bacterial therapies, the method comprising administering an
effective amount of a compound of Formula (VII): ##STR00273## or a
pharmaceutically acceptable salt thereof; wherein R.sub.13 is H,
Boc, acetyl, Fmoc, ##STR00274## R.sub.14 is H or C.sub.1-C.sub.6
alkyl; R.sub.15 is H or C.sub.1-C.sub.6 alkyl; R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; R.sub.8 is H, Boc, acetyl, Fmoc, or ##STR00275## R.sub.7
is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or
C.sub.2-C.sub.6 alkynyl; R.sub.9 is H or C.sub.1-C.sub.6 alkyl; and
R.sub.10 is H or C.sub.1-C.sub.6 alkyl
139. A process to prepare a compound of Formula (X), the process
comprises: ##STR00276## a. alkylating a compound of formula,
##STR00277## in the presence of a base and an alkylating agent; and
b. hydrolyzing the product from step a. to afford the compound of
Formula (X); wherein R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; and R.sub.18
is C.sub.1-C.sub.6 alkyl.
140. A process to prepare a compound of Formula (XI), ##STR00278##
the process comprises: a. alkylating a compound of formula,
##STR00279## in the presence of a base and an alkylating agent; and
b. hydrolyzing the product from step a. to afford the compound of
Formula (XI); wherein R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; and R.sub.18
is C.sub.1-C.sub.6 alkyl.
141. The process of claim 139 or 140, wherein the base in step a.
is an alkyllithum, a lithium bis(trialkylsilyl)amide, a sodium
bis(trialkylsilyl)amide, a potassium bis(trialkylsilyl)amide, a
lithium dialkylamide, a lithium alkoxide, a sodium alkoxide, or a
potassium alkoxide.
142. The process of claim 141, wherein the base is n-butyllithium,
lithium amide, potassium amide, sodium amide, lithium
bis(trimethylsilyl)amide, lithium diisopropylamide, potassium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide,
potassium tert-butoxide, or sodium tert-butoxide.
143. The process of claim 141, wherein the base is lithium
bis(trimethylsilyl)amide, lithium diisopropylamide, potassium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide.
144. The process of claim 141, wherein the base is sodium
bis(trimethylsilyl)amide.
145. The process of claim 139 or 140, wherein the alkylating agent
in step a. is an alkyl halide, a dialkyl sulfate, a dialkyl
carbonate, or an alkyl triflate.
146. The process of claim 145, wherein the alkylating agent is
methyl iodide, dimethyl sulfate, dimethyl carbonate, or methyl
triflate.
147. The process of claim 145, wherein the alkylating agent is
methyl iodide.
148. The process of claim 139 or 140, wherein the hydrolysis in
step b. is conducted under basic conditions.
149. The process of claim 148, wherein the basic conditions include
sodium hydroxide, lithium hydroxide, potassium hydroxide, sodium
hydroperoxide, lithium hydroperoxide, or potassium
hydroperoxide.
150. The process of claim 148, wherein the basic conditions include
lithium hydroperoxide.
Description
RELATED APPLICATIONS
[0001] The present application claims priority under 35 U.S.C.
.sctn. 119(e) to U.S. provisional application No. 62/879,246, filed
Jul. 26, 2019, which is herein incorporated by reference in its
entirety.
BACKGROUND
[0003] Quorum sensing (QS) is an intercellular communication
process adopted by a number of bacteria to regulate diverse
physiological activities. This process involves the production and
release of diffusible extracellular signaling molecules named
autoinducers (AIs), which would accumulate with increasing
bacterial population density [Miller, M. B.; Bassler, B. L., Annu.
Rev. Microbiol. 2001, 55, 165-199; Rutherford, S. T.; Bassler, B.
L., Cold Spring Harb Perspect. Med. 2012, 2 (11), a012427;
Galloway, W.; Hodgkinson, J.; Bovvden, S.; Welch, M.; Spring, D.,
Trends Microbiol. 2012, 20 (9), 449-458]. QS plays a pivotal role
in regulating bacterial pathogenesis. For example, QS modulates the
production of virulence factors such as pyocyanin and elastase in
Pseudomonas aeruginosa during bacterial growth and infection
[Jiang, Q.; Chen, J.; Yang, C.; Yin, Y.; Yao, K., BioMed. Res. Int.
2019, 1-15]. Thus, QS signaling pathway is an attractive target for
the development of antimicrobial therapeutic agents. P. aeruginosa
is a Gram-negative opportunistic pathogen that can cause serious
infections such as cystic fibrosis (CF) in lung and microbial
keratitis (MK) during contact lens wear [Sadikot, R.; Blackwell,
T.; Christman, J.; Prince, A., Am. J. Respir. Crit. Care Med. 2005,
171 (11), 1209-1223; Willcox, M. D., Optom. Vis. Sci. 2007, 84 (4),
273-278]. The AIs that control QS signaling pathway in P.
aeruginosa include two acylhomoserine lactones (AHLs,
##STR00001##
with varying alkyl chain lengths and oxidation states at C-3 and a
group of quinolone compounds (Pseudomonas quinolone signal, PQS).
They can diffuse freely across cell membranes and bind
intracellularly with corresponding receptor proteins (R proteins)
[Kwan, J. C.; Meickle, T.; Ladwa, D.; Teplitski, M.; Paul, V.;
Luesch, H., Mol. Biosyst. 2011, 7 (4), 1205-1216]. These signaling
systems form a complex hierarchical quorum sensing network, where
the Las system is considered to be the apex of the hierarchy
[Galloway, W. R.; Hodgkinson, J. T.; Bowden, S. D.; Welch, M.;
Spring, D. R., Chem. Rev. 2011, 111 (1), 28-67]. Therefore, the
LasR receptor has been usually considered to be the target for
antagonist and agonist development in P. aeruginosa [Galloway, W.
R.; Hodgkinson, J. T.; Bowden, S. D.; Welch, M.; Spring, D. R.,
Chem. Rev. 2011, 111 (1), 28-67; Hodgkinson, J. T.; Welch, M.;
Spring, D. R., ACS Chem. Biol. 2007, 2 (11), 715-717]. For example,
a synthetic non-native AHL, QSI-1, was demonstrated to be a potent
LasR antagonist [Geske, G. D.; Wezeman, R. J.; Siegel, A. P.;
Blackwell, H. E., J. Am. Chem. Soc. 2005, 127 (37), 12762-12763].
The structurally unrelated mimic of AHL, TP-1, is a highly
selective superagonist of the LasR quorum sensing system; while its
derivative TP-5 turned into a moderate QS antagonist [Muh, U.;
Hare, B. J.; Duerkop, B. A.; Schuster, M.; Hanzelka, B. L.; Heim,
R.; Olson, E. R.; Greenberg, E. P., Proc. Natl. Acad. Sci. U.S.A
2006, 103 (45), 16948-16952; Zou, Y.; Nair, S. K., Chem. Biol.
2009, 16 (9), 961-970]. Moreover, there is evidence showing that
N-octanoyl homoserine lactone can be produced in the cyanobacterium
culture of Gloeothece PCC6909 and its accumulation corresponds to a
characteristic pattern of autoinduction [Sharif, D. I.; Gallon, J.;
Smith, C. J.; Dudley, E., ISME J 2008, 2 (12), 1171-1182]. In
addition to antimicrobial (e.g., antibacterial) applications,
modulation of QS has also been shown to attenuate NF.kappa.B gene
expression and activity [Kravchenko, V. et al., Science 2008, 321,
259-263]. Additionally, modulation of QS has been demonstrated to
synergize with TRAIL to induce cancer cell death. TRAIL (tumor
necrosis factor-related apoptosis-inducing ligand), also known as
Apo-2L and TNFSF10, is a TNF family cytokine that can induce cell
apoptosis and cause programmed cell death by binding to the death
receptors DR4 (TRAIL-RI) and DR5 (TRAIL-RII) (von Karstedt, et al.
Nat Rev Cancer 2017, 17 (6), 352-366; Srivastava, at al. Neoplasia
2001, 3 (6), 535-46). Moreover, there are studies showing that
TRAIL can selectively cause cancer cell death without detrimental
effects on normal cells (Srivastava, at al. Neoplasia 2001, 3 (6),
535-46; French, et al. The TRAIL to selective tumor death. Nature
Medicine 1999, 5 (2), 146-147). This selectivity has made TRAIL a
promising candidate for cancer therapy (Trivedi, et al. Front Oncol
2015, 5, 69) and stimulated intensive studies towards the
development of therapeutic agents targeting the TRAIL signaling
pathway (von Karstedt, et al. Nat Rev Cancer 2017, 17 (6),
352-366), including recombinant TRAIL proteins and monoclonal
antibody agonists specific for DR4 (TRAIL-RI) and DR5 (TRAIL-RII)
(Amarante-Mendes, et al. Pharmacology & Therapeutics 2015, 155,
117-131). However, these agents did not achieve satisfying
anticancer activities in clinical investigations (Legler, et al.
Cell Death Dis 2018, 9 (5), 445. because many cancer cells have
developed resistance toward TRAIL, thus compromising the efficacy
of TRAIL-therapy (Trivedi, et al. Front Oncol 2015, 5, 69;
Kravchenko, et al. ACS Chem Biol 2013, 8 (6), 1117-20).
[0004] Thus, modulation of QS may be a useful approach to treat
cancer and/or increasing the effectiveness of existing anticancer
therapies.
[0005] Marine cyanobacteria have been a valuable source for the
discovery of biologically active and structurally unique natural
products including peptides, polyketides and hybrid of
peptide-polyketides. It is noteworthy that marine cyanobacterial
also produce various AHL-dependent QS inhibitors [Tang, K.; Zhang,
X. H., Mar. Drugs. 2014, 12 (6), 3245-3282]. For instance,
lyngbyoic acid,
##STR00002##
a small cyclopropane-containing fatty acid, was isolated from
Lyngbya cf. majuscula and proved to strongly inhibit the activity
of LasR [Kwan, J. C.; Meickle, T.; Ladwa, D.; Teplitski, M.; Paul,
V.; Luesch, H., Mol. Biosyst. 2011, 7 (4), 1205-1216]. Moreover,
pitinoic acid A,
##STR00003##
was also reported to be a P. aeruginosa quorum sensing inhibitor
[Montaser, R.; Paul, V. J.; Luesch, H., Org. Lett. 2013, 15 (16),
4050-4053]. In addition to these QS inhibitors, herein is reported
the isolation, total synthesis, and QS modulation activity of a
series of novel compounds.
BRIEF SUMMARY OF THE INVENTION
[0006] The invention is directed towards compounds (e.g., Formulae
(I)-(IX)), their mechanism of action, and methods of modulating
quorum sensing signaling, and methods of treating diseases and
disorders using the compounds described herein (e.g., Formulae
(I)-(IX)). In another aspect, the disease or disorder is cancer. In
another aspect, the disease or disorder is a bacterial
infection.
[0007] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00004##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0008] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0008] ##STR00005## [0009] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0010] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0011] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0012]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0012] ##STR00006## [0013] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0014] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0015] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl.
[0016] In another aspect, the invention is directed to:
##STR00007## ##STR00008##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0017] In another aspect, the invention is directed to:
##STR00009## ##STR00010## ##STR00011##
[0018] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof.
[0019] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00012##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0020] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0020] ##STR00013## [0021] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0022] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0023] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0024]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0024] ##STR00014## [0025] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0026] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0027] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl; [0028] wherein the compound is not:
##STR00015## ##STR00016##
[0028] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof.
[0029] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00017##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0030] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0030] ##STR00018## [0031] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0032] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0033] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0034]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0034] ##STR00019## [0035] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0036] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0037] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl; [0038] wherein the compound is not:
##STR00020##
[0039] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof.
[0040] In another aspect, the invention provides a pharmaceutical
composition comprising a compound described herein (e.g., Formulae
(I)-(IX)), or a pharmaceutically acceptable salt, hydrate, solvate,
or prodrug thereof, and a pharmaceutically acceptable carrier. In
another aspect, the pharmaceutical composition described herein
further comprises an additional agent. In another aspect, the
additional agent is an anti-cancer agent. In another aspect, the
additional agent is an anti-bacterial agent.
[0041] In another aspect, the invention provides a method of
modulating quorum sensing signaling, the method comprising
administering an effective amount of a compound described herein
(e.g., Formulae (I)-(IX)), or a pharmaceutically acceptable salt,
hydrate, solvate, or prodrug thereof. In another aspect, the
modulation is activation. In another aspect, the modulation is
inhibition. In another aspect, the compound is administered in
vitro. In another aspect, the compound is administered in vivo. In
another aspect, the method further comprises administering the
compound to a subject.
[0042] In another aspect, the invention provides a method of
inhibiting bacterial growth, the method comprising administering an
effective amount of a compound described herein (e.g., Formulae
(I)-(IX)), or a pharmaceutically acceptable salt, hydrate, solvate,
or prodrug thereof. In another aspect, the compound is administered
in vitro. In another aspect, the compound is administered in vivo.
In another aspect, the method further comprises administering the
compound to a subject.
[0043] In another aspect, the invention provides a method of
treating a disease or disorder in a subject in need thereof, the
method comprising administering an effective amount of a compound
described herein (e.g., Formulae (I)-(IX)), or a pharmaceutically
acceptable salt, hydrate, solvate, or prodrug thereof. In another
aspect, the disease is cancer. In another aspect, the disease is a
bacterial infection. In another aspect, the subject is a mammal. In
another aspect, the subject is a human.
[0044] In another aspect, the invention provides a method of
increasing the effectiveness of anti-cancer therapy in a subject
currently being administered one or more anti-cancer therapies, the
method comprising administering an effective amount of a compound
described herein (e.g., Formulae (I)-(IX)), or a pharmaceutically
acceptable salt, hydrate, solvate, or prodrug thereof. In certain
embodiments, the anti-cancer therapy is TRAIL. In certain
embodiments, the method increases the effectiveness of TRAIL. In
another aspect, the subject is a mammal. In another aspect, the
subject is a human.
[0045] In another aspect, the invention provides a method of
increasing the effectiveness of anti-bacterial therapy in a subject
currently being administered one or more anti-bacterial therapies,
the method comprising administering an effective amount of a
compound described herein (e.g., Formulae (I)-(IX)), or a
pharmaceutically acceptable salt, hydrate, solvate, or prodrug
thereof. In another aspect, the subject is a mammal. In another
aspect, the subject is a human.
[0046] In another aspect, the invention provides a process to
prepare a compound of Formula (X) and/or Formula (XI).
BRIEF DESCRIPTION OF THE DRAWINGS
[0047] The present invention is further described below with
reference to the following non-limiting examples and with reference
to the following figures, in which:
[0048] FIG. 1. depicts the structures of doscadenamides A-J and
doscadenamides S4-S15.
[0049] FIG. 2. depicts the .sup.1H NMR spectrum of doscadenamide A
(1a) in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0050] FIG. 3. depicts the .sup.13C NMR spectrum of doscadenamide A
(1a) in CDCl.sub.3 (150 MHz) at 27.degree. C.
[0051] FIG. 4. depicts the COSY spectrum of doscadenamide A (1a) in
CDCl.sub.3 (600 MHz) at 27.degree. C.
[0052] FIG. 5. depicts the TOCSY spectrum of doscadenamide A (1a)
in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0053] FIG. 6. depicts the HSQC spectrum of doscadenamide A (1a) in
CDCl.sub.3 (600 MHz) at 27.degree. C.
[0054] FIG. 7. depicts the HMBC spectrum of doscadenamide A (1a) in
CDCl.sub.3 (600 MHz) at 27.degree. C.
[0055] FIG. 8. depicts the NOESY spectrum of doscadenamide A (1a)
in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0056] FIG. 9. depicts the .DELTA..delta. calculation of the
.alpha.-methyl in the side chain of 1a.
[0057] FIG. 10. depicts the COSY spectrum of the (S)-PGME
derivative in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0058] FIG. 11. depicts the COSY spectrum of the (R)-PGME
derivative in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0059] FIG. 12. depicts the regional .sup.1H NMR spectra comparison
of the isolated natural product doscadenamide A and synthetic
diastereomers 1a, 1b, 1c and 1d (bottom to top, maroon, olive,
green, navy and purple, respectively) in CDCl.sub.3 (600 MHz) at
27.degree. C., with structures of the synthetic diastereomers 1a,
1b, 1c and 1d shown on the left.
[0060] FIG. 13. depicts the .sup.1H NMR spectra comparison of
natural product doscadenamide A (1a, maroon) with the four
synthetic diastereomers 1a (olive), 1b (green), 1c (navy) and 1d
(purple) in CDCl.sub.3 (600 MHz) at 27.degree. C.
[0061] FIG. 14. depicts the .sup.13C NMR spectra comparison of
natural product doscadenamide A (1a, maroon) with the four
synthetic diastereomers 1a (olive), 1b (green), 1c (navy) and 1d
(purple) in CDCl.sub.3 (150 MHz) at 27.degree. C.
[0062] FIG. 15. depicts activating activity of doscadenamide A (1a)
in pSB1075, the lasR-luxCDABE reporter constructs expressed in E.
coli. The bacterial cultures were treated with 1a in a
dose-response manner and solvent control at 37.degree. C. for 6 h
before fluorescence was measured. Results are expressed as fold
activation compared to solvent control. Data are presented as
mean.+-.SD, **P<0.01, ****P<0.0001, compared to solvent
control using one-way ANOVA (n=3).
[0063] FIG. 16. depicts activating activity of doscadenamide A (1a)
in related reporter pTIM5319, which lacks a functional AHL-binding
domain. The bacterial cultures were treated with 1a in a
dose-response manner and solvent control at 37.degree. C. for 6 h
before fluorescence was measured. Results are expressed as fold
activation compared to solvent control. Data are presented as
mean.+-.SD, **P<0.01, ****P<0.0001, compared to solvent
control using one-way ANOVA (n=3).
[0064] FIG. 17. depicts the effect of doscadenamide A (1a) and its
related diastereomers 1b, 1c, 1d, as well as positive control
3-oxo-C12-HSL (C12) at 10 .mu.M, on the production of pyocyanin in
wild-type P. aeruginosa after 6 h shaking at 37.degree. C. Data are
presented as mean.+-.SD, **P<0.01, ****P<0.0001, compared to
solvent control using one-way ANOVA (n=3).
[0065] FIG. 18 depicts the structures of doscadenamides S1-S3.
[0066] FIGS. 19A-C depict QS activity in different systems. FIG.
19A is a graph showing activity of doscadenamide A (1a) in E. coli
reporter gene assay using pSB1075 (wild-type) and point mutants
(Y56F, W60F and D73N). FIG. 19B is a graph showing normalized
pyocyanin production in P. aeruginosa mutant PAO-JP1 after
treatment with doscadenamide A (1a) and its structural analogs for
6 h at 100 .mu.M. C12 was used as the positive control. FIG. 19C is
a graph showing measurement of QS activating activity of
doscadenamide A (1a) and its structural analogs in V. harveyi after
8 h treatment, presented by .DELTA.LUM [.DELTA.LUM=LUM
(doscadenamide analog)-LUM (DMSO)] normalized by cell viability
measured with OD600. AI-1 is a reported quorum sensing autoinducer
and was used as the positive control.
[0067] FIGS. 20A-E depict that doscadenamide A (1a) and its
structural analogs sensitize breast cancer MDA-MB-231 cells in
combination with TRAIL. FIGS. 20A-C are graphs showing
dose-response analysis of TRAIL on MDA-MB-231 cells. MDA-MB-231
cell viability after 3 h pretreatment with DMSO (solvent control),
C12 as well as doscadenamide A (1a) and its structural analogs, B)
50 .mu.M and C) 25 .mu.M followed by combined TRAIL (20 ng/mL)
treatment for 24 h. FIG. 20D is a chart showing A Bliss
independence calculations for MDA-MB-231 cells cotreated with TRAIL
(20 ng/mL) and doscadenamide A (1a) and its structural analogs.
MDA-MB-231 cells were treated with doscadenamide A (1a) and its
structural analogs for 3 h, followed by treatment with TRAIL (20
ng/mL) for 24 h, C12 was included as positive control. "A Bliss
independence" is the difference between observed growth inhibition
and Bliss expectation. Values greater than zero represent a
synergistic response, represented as red in the figure. Bliss
expectation is C=(A+B)-(A.times.B), where A and B are the growth
inhibition fractions of two compounds at a given dose. Cell
viability was quantified using MTT assay. FIG. 20E is a western
blot analysis of protein extracts from breast cancer MDA-MB-231
cells after treatment with TRAIL (20 ng/mL), doscadenamide A (1a)
and its analogs (50 .mu.M) and their combination as shown in the
figure.
DETAILED DESCRIPTION
Definitions
[0068] In order that the invention may be more readily understood,
certain terms are first defined here for convenience.
[0069] As used herein, the term "treating" a disorder encompasses
ameliorating, mitigating and/or managing the disorder and/or
conditions that may cause the disorder. The terms "treating" and
"treatment" refer to a method of alleviating or abating a disease
and/or its attendant symptoms. In accordance with the present
invention, "treating" includes blocking, inhibiting, attenuating,
modulating, reversing the effects of and reducing the occurrence of
e.g., the harmful effects of a disorder.
[0070] As used herein, "inhibiting" encompasses reducing and
halting progression.
[0071] The term "modulate" refers to increases or decreases in the
activity of a cell in response to exposure to a compound of the
invention.
[0072] The terms "isolated," "purified," or "biologically pure"
refer to material that is substantially or essentially free from
components that normally accompany it as found in its native state.
Purity and homogeneity are typically determined using analytical
chemistry techniques such as polyacrylamide gel electrophoresis or
high performance liquid chromatography. Particularly, in
embodiments the compound is at least 85% pure, more preferably at
least 90% pure, more preferably at least 95% pure, and most
preferably at least 99% pure.
[0073] The terms "polypeptide," "peptide" and "protein" are used
interchangeably herein to refer to a polymer of amino acid
residues. The terms apply to amino acid polymers in which one or
more amino acid residue is an artificial chemical mimetic of a
corresponding naturally occurring amino acid, as well as to
naturally occurring amino acid polymers and non-naturally occurring
amino acid polymer.
[0074] A "peptide" is a sequence of at least two amino acids.
Peptides can consist of short as well as long amino acid sequences,
including proteins.
[0075] The term "amino acid" refers to naturally occurring and
synthetic amino acids, as well as amino acid analogs and amino acid
mimetics that function in a manner similar to the naturally
occurring amino acids. Naturally occurring amino acids are those
encoded by the genetic code, as well as those amino acids that are
later modified, e.g., hydroxyproline, .gamma.-carboxyglutamate, and
O-phosphoserine. Amino acid analogs refers to compounds that have
the same basic chemical structure as a naturally occurring amino
acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl
group, an amino group, and an R group, e.g., homoserine,
norleucine, methionine sulfoxide, methionine methyl sulfonium. Such
analogs have modified R groups (e.g., norleucine) or modified
peptide backbones, but retain the same basic chemical structure as
a naturally occurring amino acid. Amino acid mimetics refers to
chemical compounds that have a structure that is different from the
general chemical structure of an amino acid, but that functions in
a manner similar to a naturally occurring amino acid.
[0076] The term "protein" refers to series of amino acid residues
connected one to the other by peptide bonds between the alpha-amino
and carboxy groups of adjacent residues.
[0077] Amino acids may be referred to herein by either their
commonly known three letter symbols or by the one-letter symbols
recommended by the IUPAC-IUB Biochemical Nomenclature
Commission.
[0078] As to amino acid sequences, one of skill will recognize that
individual substitutions, deletions or additions to a peptide,
polypeptide, or protein sequence which alters, adds or deletes a
single amino acid or a small percentage of amino acids in the
encoded sequence is a "conservatively modified variant" where the
alteration results in the substitution of an amino acid with a
chemically similar amino acid. Conservative substitution tables
providing functionally similar amino acids are well known in the
art.
[0079] Macromolecular structures such as polypeptide structures can
be described in terms of various levels of organization. For a
general discussion of this organization, see, e.g., Alberts et al.,
Molecular Biology of the Cell (3rd ed., 1994) and Cantor and
Schimmel, Biophysical Chemistry Part I. The Conformation of
Biological Macromolecules (1980). "Primary structure" refers to the
amino acid sequence of a particular peptide. "Secondary structure"
refers to locally ordered, three dimensional structures within a
polypeptide. These structures are commonly known as domains.
Domains are portions of a polypeptide that form a compact unit of
the polypeptide and are typically 50 to 350 amino acids long.
Typical domains are made up of sections of lesser organization such
as stretches of .beta.-sheet and .alpha.-helices. "Tertiary
structure" refers to the complete three-dimensional structure of a
polypeptide monomer. "Quaternary structure" refers to the
three-dimensional structure formed by the noncovalent association
of independent tertiary units. Anisotropic terms are also known as
energy terms.
[0080] The term "administration" or "administering" includes routes
of introducing the compound(s) to a subject to perform their
intended function. Examples of routes of administration which can
be used include injection (subcutaneous, intravenous, parenterally,
intraperitoneally, intrathecal), topical, oral, inhalation, rectal
and transdermal.
[0081] The term "effective amount" includes an amount effective, at
dosages and for periods of time necessary, to achieve the desired
result. An effective amount of compound may vary according to
factors such as the disease state, age, and weight of the subject,
and the ability of the compound to elicit a desired response in the
subject. Dosage regimens may be adjusted to provide the optimum
therapeutic response. An effective amount is also one in which any
toxic or detrimental effects (e.g., side effects) of the elastase
inhibitor compound are outweighed by the therapeutically beneficial
effects.
[0082] The phrases "systemic administration," "administered
systemically", "peripheral administration" and "administered
peripherally" as used herein mean the administration of a
compound(s), drug or other material, such that it enters the
patient's system and, thus, is subject to metabolism and other like
processes.
[0083] The term "therapeutically effective amount" refers to that
amount of the compound being administered sufficient to prevent
development of or alleviate to some extent one or more of the
symptoms of the condition or disorder being treated.
[0084] A therapeutically effective amount of compound (i.e., an
effective dosage) may range from about 0.005 .mu.g/kg to about 200
mg/kg, preferably about 0.1 mg/kg to about 200 mg/kg, more
preferably about 10 mg/kg to about 100 mg/kg of body weight. In
other embodiments, the therapeutically effect amount may range from
about 1.0 pM to about 500 nM. The skilled artisan will appreciate
that certain factors may influence the dosage required to
effectively treat a subject, including but not limited to the
severity of the disease or disorder, previous treatments, the
general health and/or age of the subject, and other diseases
present. Moreover, treatment of a subject with a therapeutically
effective amount of a compound can include a single treatment or,
preferably, can include a series of treatments. In one example, a
subject is treated with a compound in the range of between about
0.005 .mu.g/kg to about 200 mg/kg of body weight, one time per week
for between about 1 to 10 weeks, preferably between 2 to 8 weeks,
more preferably between about 3 to 7 weeks, and even more
preferably for about 4, 5, or 6 weeks. It will also be appreciated
that the effective dosage of a compound used for treatment may
increase or decrease over the course of a particular treatment.
[0085] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0086] The term "diastereomers" refers to stereoisomers with two or
more centers of dissymmetry and whose molecules are not mirror
images of one another.
[0087] The term "enantiomers" refers to two stereoisomers of a
compound which are non-superimposable mirror images of one another.
An equimolar mixture of two enantiomers is called a "racemic
mixture" or a "racemate."
[0088] The term "isomers" or "stereoisomers" refers to compounds
which have identical chemical constitution, but differ with regard
to the arrangement of the atoms or groups in space.
[0089] The term "prodrug" includes compounds with moieties which
can be metabolized in vivo. Generally, the prodrugs are metabolized
in vivo by esterases or by other mechanisms to active drugs.
Examples of prodrugs and their uses are well known in the art (See,
e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci.
66:1-19). The prodrugs can be prepared in situ during the final
isolation and purification of the compounds, or by separately
reacting the purified compound in its free acid form or hydroxyl
with a suitable esterifying agent. Hydroxyl groups can be converted
into esters via treatment with a carboxylic acid. Examples of
prodrug moieties include substituted and unsubstituted, branch or
unbranched lower alkyl ester moieties, (e.g., propionoic acid
esters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl
esters (e.g., dimethylaminoethyl ester), acylamino lower alkyl
esters (e.g., acetyloxymethyl ester), acyloxy lower alkyl esters
(e.g., pivaloyloxymethyl ester), aryl esters (phenyl ester),
aryl-lower alkyl esters (e.g., benzyl ester), substituted (e.g.,
with methyl, halo, or methoxy substituents) aryl and aryl-lower
alkyl esters, amides, lower-alkyl amides, di-lower alkyl amides,
and hydroxy amides. Preferred prodrug moieties are propionoic acid
esters and acyl esters. Prodrugs which are converted to active
forms through other mechanisms in vivo are also included.
[0090] The term "subject" refers to animals such as mammals,
including, but not limited to, primates (e.g., humans), cows,
sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like.
In certain embodiments, the subject is a human.
[0091] Furthermore, the compounds of the invention include olefins
having either geometry: "Z" refers to what is referred to as a
"cis" (same side) conformation whereas "E" refers to what is
referred to as a "trans" (opposite side) conformation. With respect
to the nomenclature of a chiral center, the terms "d" and "1"
configuration are as defined by the IUPAC Recommendations. As to
the use of the terms, diastereomer, racemate, epimer and
enantiomer, these will be used in their normal context to describe
the stereochemistry of preparations.
[0092] As used herein, the term "alkyl" refers to a
straight-chained or branched hydrocarbon group containing 1 to 12
carbon atoms. The term "lower alkyl" refers to a C.sub.1-C.sub.6
alkyl chain. Examples of alkyl groups include methyl, ethyl,
n-propyl, isopropyl, tert-butyl, and n-pentyl. Alkyl groups may be
optionally substituted with one or more substituents.
[0093] The term "alkenyl" refers to an unsaturated hydrocarbon
chain that may be a straight chain or branched chain, containing 2
to 12 carbon atoms and at least one carbon-carbon double bond.
Alkenyl groups may be optionally substituted with one or more
substituents.
[0094] The term "alkynyl" refers to an unsaturated hydrocarbon
chain that may be a straight chain or branched chain, containing
the 2 to 12 carbon atoms and at least one carbon-carbon triple
bond. Alkynyl groups may be optionally substituted with one or more
substituents.
[0095] The sp.sup.2 or sp carbons of an alkenyl group and an
alkynyl group, respectively, may optionally be the point of
attachment of the alkenyl or alkynyl groups.
[0096] The term "alkoxy" refers to an --O-alkyl radical.
[0097] As used herein, the term "halogen", "hal" or "halo" means
--F, --Cl, --Br or --I.
[0098] The term "cycloalkyl" refers to a hydrocarbon 3-8 membered
monocyclic or 7-14 membered bicyclic ring system having at least
one saturated ring or having at least one non-aromatic ring,
wherein the non-aromatic ring may have some degree of unsaturation.
Cycloalkyl groups may be optionally substituted with one or more
substituents. In one embodiment, 0, 1, 2, 3, or 4 atoms of each
ring of a cycloalkyl group may be substituted by a substituent.
Representative examples of cycloalkyl group include cyclopropyl,
cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl, cyclopentenyl,
cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like.
[0099] The term "aryl" refers to a hydrocarbon monocyclic, bicyclic
or tricyclic aromatic ring system. Aryl groups may be optionally
substituted with one or more substituents. In one embodiment, 0, 1,
2, 3, 4, 5 or 6 atoms of each ring of an aryl group may be
substituted by a substituent. Examples of aryl groups include
phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, and
the like.
[0100] The term "heteroaryl" refers to an aromatic 5-8 membered
monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic
ring system having 1-4 ring heteroatoms if monocyclic, 1-6
heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said
heteroatoms selected from O, N, or S, and the remainder ring atoms
being carbon (with appropriate hydrogen atoms unless otherwise
indicated). Heteroaryl groups may be optionally substituted with
one or more substituents. In one embodiment, 0, 1, 2, 3, or 4 atoms
of each ring of a heteroaryl group may be substituted by a
substituent. Examples of heteroaryl groups include pyridyl,
furanyl, thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl
thiazolyl, isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl,
pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, isoquinolinyl,
indazolyl, and the like.
[0101] The term "heterocycloalkyl" refers to a nonaromatic 3-8
membered monocyclic, 7-12 membered bicyclic, or 10-14 membered
tricyclic ring system comprising 1-3 heteroatoms if monocyclic, 1-6
heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said
heteroatoms selected from O, N, S, B, P or Si, wherein the
nonaromatic ring system is completely saturated. Heterocycloalkyl
groups may be optionally substituted with one or more substituents.
In one embodiment, 0, 1, 2, 3, or 4 atoms of each ring of a
heterocycloalkyl group may be substituted by a substituent.
Representative heterocycloalkyl groups include piperidinyl,
piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl,
1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl, thiirenyl, and
the like.
[0102] The term "alkylamino" refers to an amino substituent which
is further substituted with one or two alkyl groups. The term
"aminoalkyl" refers to an alkyl substituent which is further
substituted with one or more amino groups. The term "hydroxyalkyl"
or "hydroxylalkyl" refers to an alkyl substituent which is further
substituted with one or more hydroxyl groups. The alkyl or aryl
portion of alkylamino, aminoalkyl, mercaptoalkyl, hydroxyalkyl,
mercaptoalkoxy, sulfonylalkyl, sulfonylaryl, alkylcarbonyl, and
alkylcarbonylalkyl may be optionally substituted with one or more
substituents.
[0103] Acids and bases useful in the methods herein are known in
the art. Acid catalysts are any acidic chemical, which can be
inorganic (e.g., hydrochloric, hydrobromic, sulfuric, nitric acids,
aluminum trichloride) or organic (e.g., camphorsulfonic acid,
p-toluenesulfonic acid, acetic acid, ytterbium triflate) in nature.
Acids are useful in either catalytic or stoichiometric amounts to
facilitate chemical reactions. Bases are any basic chemical, which
can be inorganic (e.g., sodium bicarbonate, potassium hydroxide) or
organic (e.g., triethylamine, pyridine) in nature. Bases are useful
in either catalytic or stoichiometric amounts to facilitate
chemical reactions.
[0104] Alkylating agents are any reagent that is capable of
effecting the alkylation of the functional group at issue (e.g.,
oxygen atom of an alcohol, nitrogen atom of an amino group).
Alkylating agents are known in the art, including in the references
cited herein, and include alkyl halides (e.g., methyl iodide,
benzyl bromide or chloride), alkyl sulfates (e.g., methyl sulfate),
or other alkyl group-leaving group combinations known in the art.
Leaving groups are any stable species that can detach from a
molecule during a reaction (e.g., elimination reaction,
substitution reaction) and are known in the art, including in the
references cited herein, and include halides (e.g., I--, Cl--,
Br--, F--), hydroxy, alkoxy (e.g., --OMe, --O-t-Bu), acyloxy anions
(e.g., --OAc, --OC(O)CF.sub.3), sulfonates (e.g., mesyl, tosyl),
acetamides (e.g., --NHC(O)Me), carbamates (e.g., N(Me)C(O)Ot-Bu),
phosphonates (e.g., --OP(O) (OEt).sub.2), water or alcohols (protic
conditions), and the like.
[0105] In certain embodiments, substituents on any group (such as,
for example, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, cycloalkyl, heterocycloalkyl) can be at any atom of
that group, wherein any group that can be substituted (such as, for
example, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroaralkyl, cycloalkyl, heterocycloalkyl) can be optionally
substituted with one or more substituents (which may be the same or
different), each replacing a hydrogen atom. Examples of suitable
substituents include, but are not limited to alkyl, alkenyl,
alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl,
aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy,
aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl,
formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl,
alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy,
heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl,
amino, aminoalkyl, dialkylamino, alkylcarbonylamino,
alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino,
diarylamino, alkylcarbonyl, or arylamino-substituted aryl;
arylalkylamino, aralkylaminocarbonyl, amido, alkylaminosulfonyl,
arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino,
arylsulfonylamino, imino, carbamido, carbamyl, thioureido,
thiocyanato, sulfoamido, sulfonylalkyl, sulfonylaryl, or
mercaptoalkoxy.
Compounds of the Invention
[0106] Compounds delineated herein (e.g., Formulae (I)-(XI))
include salt, hydrate and solvates thereof. They include all
compounds delineated in schemes herein, whether intermediate or
final compounds in a process.
[0107] Compounds of the invention can be obtained from natural
sources or made or modified made by means known in the art of
organic synthesis. Methods for optimizing reaction conditions, if
necessary, minimizing competing by-products, are known in the art.
Reaction optimization and scale-up may advantageously utilize
high-speed parallel synthesis equipment and computer-controlled
microreactors (e.g., Design And Optimization in Organic Synthesis,
2.sup.nd Edition, Carlson R, Ed, 2005; Elsevier Science Ltd.;
Jahnisch, K et al, Angew. Chem. Int. Ed. Engl. 2004 43: 406; and
references therein). Additional reaction schemes and protocols may
be determined by the skilled artisan by use of commercially
available structure-searchable database software, for instance,
SciFinder.RTM. (CAS division of the American Chemical Society) and
CrossFire Beilstein.RTM. (Elsevier MDL), or by appropriate keyword
searching using an internet search engine such as Google.RTM. or
keyword databases such as the US Patent and Trademark Office text
database.
[0108] The compounds herein may also contain linkages (e.g.,
carbon-carbon bonds) wherein bond rotation is restricted about that
particular linkage, e.g. restriction resulting from the presence of
a ring or double bond. Accordingly, all cis/trans and E/Z isomers
are expressly included in the present invention. The compounds
herein may also be represented in multiple tautomeric forms, in
such instances, the invention expressly includes all tautomeric
forms of the compounds described herein, even though only a single
tautomeric form may be represented. All such isomeric forms of such
compounds herein are expressly included in the present invention.
All crystal forms and polymorphs of the compounds described herein
are expressly included in the present invention. All hydrate and
solvate forms of the compounds described herein are expressly
included in the present invention. Also embodied are extracts and
fractions comprising compounds of the invention. The term isomers
is intended to include diastereoisomers, enantiomers, regioisomers,
structural isomers, rotational isomers, tautomers, and the like.
For compounds which contain one or more stereogenic centers, e.g.,
chiral compounds, the methods of the invention may be carried out
with an enantiomerically enriched compound, a racemate, or a
mixture of diastereomers.
[0109] Preferred enantiomerically enriched compounds have an
enantiomeric excess of 50% or more, more preferably the compound
has an enantiomeric excess of 60%, 70%, 80%, 90%, 95%, 98%, or 99%
or more. In preferred embodiments, only one enantiomer or
diastereomer of a chiral compound of the invention is administered
to cells or a subject.
[0110] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00021##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0111] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0111] ##STR00022## [0112] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0113] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0114] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0115]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0115] ##STR00023## [0116] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0117] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0118] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl.
[0119] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00024##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0120] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0120] ##STR00025## [0121] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0122] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0123] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl;
[0123] ##STR00026## [0124] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0125] R.sub.7 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; [0126] R.sub.9 is H or
C.sub.1-C.sub.6 alkyl; and [0127] R.sub.10 is H or C.sub.1-C.sub.6
alkyl; [0128] wherein the compound is not:
##STR00027##
[0129] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof.
[0130] In another aspect, the invention is directed to a compound
of Formula (IX):
##STR00028##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0131] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0131] ##STR00029## [0132] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0133] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0134] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl;
[0134] ##STR00030## [0135] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0136] R.sub.7 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; [0137] R.sub.9 is H or
C.sub.1-C.sub.6 alkyl; and [0138] R.sub.10 is H or C.sub.1-C.sub.6
alkyl; [0139] wherein the compound is not:
##STR00031##
[0139] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof. In another aspect, R.sub.8 is
##STR00032##
In another aspect, R.sub.9 is H. In another aspect, R.sub.10 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.9 is H and R.sub.10
is C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.10 is Me. In
another aspect, R.sub.10 is H. In another aspect, R.sub.9 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.10 is H and R.sub.9
is C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.9 is Me. In
another aspect, R.sub.7 is C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.7 is
##STR00033##
In another aspect, R.sub.7 is C.sub.2-C.sub.6 alkenyl. In another
aspect, R.sub.7 is
##STR00034##
In another aspect, R.sub.7 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.7 is
##STR00035##
[0140] In another aspect, R.sub.13 is
##STR00036##
In another aspect R.sub.14 is H. In another aspect, R.sub.14 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.14 is Me. In
another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.6 is
##STR00037##
In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl. In another
aspect, R.sub.6 is
##STR00038##
In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl. In another
aspect R.sub.6 is
##STR00039##
[0141] In another aspect, R.sub.13 is
##STR00040##
In another aspect R.sub.14 is H. In another aspect, R.sub.14 is H
and R.sub.15 is C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.14
is H and R.sub.15 is Me. In another aspect, R.sub.15 is H. In
another aspect, R.sub.15 is H and R.sub.14 is C.sub.1-C.sub.6
alkyl. In another aspect, R.sub.15 is H and R.sub.14 is Me. In
another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.6 is
##STR00041##
In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl. In another
aspect, R.sub.6 is
##STR00042##
In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl. In another
aspect R.sub.6 is
##STR00043##
[0142] In another aspect, the invention is directed to a compound
of Formula (I):
##STR00044##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0143] wherein R.sub.1 is H, Boc, acetyl, Fmoc, or
[0143] ##STR00045## [0144] R.sub.2 is H or C.sub.1-C.sub.6 alkyl;
[0145] R.sub.3 is H; [0146] R.sub.4 is H or C.sub.1-C.sub.6 alkyl;
[0147] R.sub.5 is H or C.sub.1-C.sub.6 alkyl; [0148] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; and [0149] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.1 is
[0149] ##STR00046## In another aspect, R.sub.2 is C.sub.1-C.sub.6
alkyl. In another aspect, R.sub.2 is Me. In another aspect, R.sub.7
is C.sub.2-C.sub.6 alkynyl. In another aspect, R.sub.7 is
##STR00047## In another aspect, R.sub.7 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.7 is
##STR00048## In another aspect, R.sub.7 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.7 is
##STR00049## In another aspect R.sub.4 is H. In another aspect,
R.sub.4 is H and R.sub.5 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.4 is H and R.sub.5 is Me. In another aspect, R.sub.5
is H. In another aspect, R.sub.5 is H and R.sub.4 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.5 is H and R.sub.4
is Me. In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl. In
another aspect, R.sub.6 is
##STR00050## In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.6 is
##STR00051## In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.6 is
##STR00052##
[0150] In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is according to Formula (II):
##STR00053##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0151] In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is according to Formula (III):
##STR00054##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0152] In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is according to Formula (IV):
##STR00055##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0153] In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is
##STR00056##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0154] In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is
##STR00057##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof. In another aspect, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is
##STR00058##
[0155] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof.
[0156] In certain embodiments, the compound of Formula (I), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is not
##STR00059##
[0157] In another aspect, the invention is directed to a compound
of Formula (V):
##STR00060##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0158] wherein R.sub.4 is H or C.sub.1-C.sub.6 alkyl;
[0159] R.sub.5 is H or C.sub.1-C.sub.6 alkyl; [0160] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0161] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0161] ##STR00061## [0162] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0163] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0164] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.8 is
[0164] ##STR00062## In another aspect, R.sub.9 is H. In another
aspect, R.sub.10 is C.sub.1-C.sub.6 alkyl. In another aspect,
R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6 alkyl. In another
aspect, wherein R.sub.10 is Me. In another aspect, R.sub.10 is H.
In another aspect, R.sub.9 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6 alkyl. In
another aspect, wherein R.sub.9 is Me. In another aspect, R.sub.7
is C.sub.2-C.sub.6 alkynyl. In another aspect, R.sub.7 is
##STR00063## In another aspect, R.sub.7 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.7 is
##STR00064##
In another aspect, R.sub.7 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.7 is
##STR00065##
In another aspect, R.sub.4 is H. In another aspect, R.sub.4 is H
and R.sub.5 is C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.4 is
H and R.sub.5 is Me. In another aspect, R.sub.5 is H. In another
aspect, R.sub.5 is H and R.sub.4 is C.sub.1-C.sub.6 alkyl. In
another aspect, R.sub.5 is H and R.sub.4 is Me. In another aspect,
R.sub.6 is C.sub.2-C.sub.6 alkynyl. In another aspect, R.sub.6
is
##STR00066## In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.6 is
##STR00067## In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.6 is
##STR00068## In another aspect the compound is
##STR00069##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0165] In another aspect, the invention is directed to a compound
of Formula (VI):
##STR00070##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0166] wherein Ru is H, Boc, acetyl, Fmoc, or
[0166] ##STR00071## [0167] R.sub.11 is C.sub.1-C.sub.6 alkyl;
[0168] R.sub.6 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; [0169] R.sub.8 is H, Boc,
acetyl, Fmoc, or
[0169] ##STR00072## [0170] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0171] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0172] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.11 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.11 is Me. In
another aspect, R.sub.8 is
[0172] ##STR00073## In another aspect, R.sub.9 is H. In another
aspect, R.sub.10 is C.sub.1-C.sub.6 alkyl. In another aspect,
R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.10 is Me. In another aspect, R.sub.10 is H. In
another aspect, R.sub.9 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6 alkyl. In
another aspect, R.sub.9 is Me. In another aspect, R.sub.7 is
C.sub.2-C.sub.6 alkynyl. In another aspect, R.sub.7 is
##STR00074## In another aspect, R.sub.7 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.7 is
##STR00075## In another aspect, R.sub.7 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.7 is
##STR00076## In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl.
In another aspect, R.sub.6 is
##STR00077## In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.6 is
##STR00078## In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.6 is
##STR00079## In another aspect, the compound is:
##STR00080##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0173] In another aspect, the compound is:
##STR00081##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0174] In another aspect, the invention is directed to a compound
of Formula (VII):
##STR00082##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0175] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0175] ##STR00083## [0176] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0177] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0178] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0179] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0179] ##STR00084## [0180] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0181] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0182] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.8 is
[0182] ##STR00085## In another aspect, R.sub.9 is H. In another
aspect, R.sub.10 is C.sub.1-C.sub.6 alkyl. In another aspect,
R.sub.9 is H and R.sub.10 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.10 is Me. In another aspect, R.sub.10 is H. In
another aspect, R.sub.9 is C.sub.1-C.sub.6 alkyl. In another
aspect, R.sub.10 is H and R.sub.9 is C.sub.1-C.sub.6 alkyl. In
another aspect, R.sub.9 is Me. In another aspect, R.sub.7 is
C.sub.2-C.sub.6 alkynyl. In another aspect, R.sub.7 is
##STR00086## In another aspect, R.sub.7 is C.sub.2-C.sub.6 alkenyl.
In another aspect, R.sub.7 is
##STR00087## In another aspect, R.sub.7 is C.sub.1-C.sub.6 alkyl.
In another aspect, R.sub.7 is
##STR00088##
[0183] In another aspect, R.sub.13 is
##STR00089##
In another aspect R.sub.14 is H. In another aspect, R.sub.14 is
C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.14 is Me. In
another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.6 is
##STR00090##
In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl. In another
aspect, R.sub.6 is
##STR00091##
In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl. In another
aspect R.sub.6 is
##STR00092##
[0184] In another aspect, R.sub.13 is
##STR00093##
In another aspect R.sub.14 is H. In another aspect, R.sub.14 is H
and R.sub.15 is C.sub.1-C.sub.6 alkyl. In another aspect, R.sub.14
is H and R.sub.15 is Me. In another aspect, R.sub.15 is H. In
another aspect, R.sub.15 is H and R.sub.14 is C.sub.1-C.sub.6
alkyl. In another aspect, R.sub.15 is H and R.sub.14 is Me. In
another aspect, R.sub.6 is C.sub.2-C.sub.6 alkynyl. In another
aspect, R.sub.6 is
##STR00094##
In another aspect, R.sub.6 is C.sub.2-C.sub.6 alkenyl. In another
aspect, R.sub.6 is
##STR00095##
In another aspect, R.sub.6 is C.sub.1-C.sub.6 alkyl. In another
aspect R.sub.6 is
##STR00096##
[0185] In another aspect, the compound of Formula (VII), or a
pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof, is
##STR00097##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof.
[0186] In another aspect, the invention is directed to a compound
of Formula (VIII):
##STR00098##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0187] wherein R.sub.16 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; and Rig is H
or C.sub.1-C.sub.6 alkyl; or [0188] R.sub.16 is H or
C.sub.1-C.sub.6 alkyl; and Rig is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl. In another
aspect, the compound is
[0188] ##STR00099## or a pharmaceutically acceptable salt, solvate,
hydrate, or prodrug thereof.
[0189] In another aspect, the invention is directed to a process to
prepare a compound of Formula (X), the process comprises:
##STR00100## [0190] a. alkylating a compound of formula,
[0190] ##STR00101## in the presence of a base and an alkylating
agent; and [0191] b. hydrolyzing the product from step a. to afford
the compound of Formula (X); [0192] wherein R.sub.7 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; and [0193] R.sub.18 is C.sub.1-C.sub.6 alkyl. In another
aspect, the compound in step a. is
[0193] ##STR00102## In another aspect, the base in step a. is an
alkyllithum, a lithium bis(trialkylsilyl)amide, a sodium
bis(trialkylsilyl)amide, a potassium bis(trialkylsilyl)amide, a
lithium dialkylamide, a lithium alkoxide, a sodium alkoxide, or a
potassium alkoxide. In another aspect, the base in step a. is
n-butyllithium, lithium amide, potassium amide, sodium amide,
lithium bis(trimethylsilyl)amide, lithium diisopropylamide,
potassium bis(trimethylsilyl)amide, sodium
bis(trimethylsilyl)amide, potassium tert-butoxide, or sodium
tert-butoxide. In another aspect, the base in step a. is lithium
bis(trimethylsilyl)amide, lithium diisopropylamide, potassium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide. In
another aspect, the base in step a. is sodium
bis(trimethylsilyl)amide. In another aspect, the alkylating agent
in step a. is an alkyl halide, a dialkyl sulfate, a dialkyl
carbonate, or an alkyl triflate. In another aspect, the alkylating
agent in step a. is methyl iodide, dimethyl sulfate, dimethyl
carbonate, or methyl triflate. In another aspect, the alkylating
agent in step a. is methyl iodide. In another aspect, the
hydrolysis in step b. is conducted under basic conditions. In
another aspect, the basic conditions include sodium hydroxide,
lithium hydroxide, potassium hydroxide, sodium hydroperoxide,
lithium hydroperoxide, or potassium hydroperoxide. In another
aspect, the basic conditions include lithium hydroperoxide.
[0194] In another aspect, the invention is directed to a process to
prepare a compound of Formula (XI),
##STR00103##
the process comprises: [0195] a. alkylating a compound of
formula,
[0195] ##STR00104## in the presence of a base and an alkylating
agent; and [0196] b. hydrolyzing the product from step a. to afford
the compound of Formula (XI); [0197] wherein R.sub.7 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; and [0198] R.sub.18 is C.sub.1-C.sub.6 alkyl. In another
aspect, the compound in step a. is
[0198] ##STR00105## In another aspect, the base in step a. is an
alkyllithum, a lithium bis(trialkylsilyl)amide, a sodium
bis(trialkylsilyl)amide, a potassium bis(trialkylsilyl)amide, a
lithium dialkylamide, a lithium alkoxide, a sodium alkoxide, or a
potassium alkoxide. In another aspect, the base in step a. is
n-butyllithium, lithium amide, potassium amide, sodium amide,
lithium bis(trimethylsilyl)amide, lithium diisopropylamide,
potassium bis(trimethylsilyl)amide, sodium
bis(trimethylsilyl)amide, potassium tert-butoxide, or sodium
tert-butoxide. In another aspect, the base in step a. is lithium
bis(trimethylsilyl)amide, lithium diisopropylamide, potassium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide. In
another aspect, the base in step a. is sodium
bis(trimethylsilyl)amide. In another aspect, the alkylating agent
in step a. is an alkyl halide, a dialkyl sulfate, a dialkyl
carbonate, or an alkyl triflate. In another aspect, the alkylating
agent in step a. is methyl iodide, dimethyl sulfate, dimethyl
carbonate, or methyl triflate. In another aspect, the alkylating
agent in step a. is methyl iodide. In another aspect, the
hydrolysis in step b. is conducted under basic conditions. In
another aspect, the basic conditions include sodium hydroxide,
lithium hydroxide, potassium hydroxide, sodium hydroperoxide,
lithium hydroperoxide, or potassium hydroperoxide. In another
aspect, the basic conditions include lithium hydroperoxide.
Methods of Treatment
[0199] In another aspect, the invention provides a method of
modulating quorum sensing signaling, the method comprising
administering an effective amount of a compound of Formula
(IX):
##STR00106##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0200] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0200] ##STR00107## [0201] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0202] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0203] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0204]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0204] ##STR00108## [0205] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0206] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0207] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the modulation is
activation. In another aspect, the modulation is inhibition. In
another aspect, the compound is administered in vitro. In another
aspect, the compound is administered in vivo. In another aspect,
the method further comprises administering the compound to a
subject.
[0208] In another aspect, the invention provides a method of
modulating quorum sensing signaling, the method comprising
administering an effective amount of a compound of Formula
(VII):
##STR00109##
[0209] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0210] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0210] ##STR00110## [0211] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0212] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0213] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0214] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0214] ##STR00111## [0215] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0216] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0217] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the modulation is
activation. In another aspect, the modulation is inhibition. In
another aspect, the compound is administered in vitro. In another
aspect, the compound is administered in vivo. In another aspect,
the method further comprises administering the compound to a
subject.
[0218] In another aspect, the invention provides a method of
inhibiting bacterial growth, the method comprising administering an
effective amount of a compound of Formula (IX):
##STR00112##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0219] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0219] ##STR00113## [0220] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0221] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0222] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0223]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0223] ##STR00114## [0224] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0225] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0226] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the compound is
administered in vitro. In another aspect, the compound is
administered in vivo. In another aspect, the method further
comprises administering the compound to a subject.
[0227] In another aspect, the invention provides a method of
inhibiting bacterial growth, the method comprising administering an
effective amount of a compound of Formula (VII):
##STR00115##
[0228] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0229] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0229] ##STR00116## [0230] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0231] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0232] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0233] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0233] ##STR00117## [0234] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0235] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0236] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the compound is
administered in vitro. In another aspect, the compound is
administered in vivo. In another aspect, the method further
comprises administering the compound to a subject.
[0237] In another aspect, the invention provides a method of
treating a bacterial infection in a subject, the method comprising
administering an effective amount of a compound of Formula
(IX):
##STR00118##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0238] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0238] ##STR00119## [0239] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0240] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0241] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0242]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0242] ##STR00120## [0243] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0244] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0245] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0246] In another aspect, the invention provides a method of
treating a bacterial infection in a subject in need thereof, the
method comprising administering an effective amount of a compound
of Formula (IX):
##STR00121##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0247] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0247] ##STR00122## [0248] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0249] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0250] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0251]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0251] ##STR00123## [0252] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0253] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0254] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0255] In another aspect, the invention provides a method of
treating a bacterial infection in a subject, the method comprising
administering an effective amount of a compound of Formula
(VII):
##STR00124##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0256] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0256] ##STR00125## [0257] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0258] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0259] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0260] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0260] ##STR00126## [0261] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0262] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0263] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0264] In another aspect, the invention provides a method of
treating a bacterial infection in a subject in need thereof, the
method comprising administering an effective amount of a compound
of Formula (VII):
##STR00127##
[0265] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0266] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0266] ##STR00128## [0267] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0268] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0269] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0270] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0270] ##STR00129## [0271] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0272] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0273] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0274] In certain embodiments, the bacterial infection is an
infection caused by Gram-positive bacteria. In certain embodiments,
the bacterial infection is an infection caused by Gram-negative
bacteria. In certain embodiments, the bacterial infection is a
Staphylococcus infection, a Bacillus infection, or an Escherichia
infection. In some embodiments, the bacterial infection is caused
by a member of Mycobacteriacae. In certain embodiments, the
bacterial infection is an infection caused by Mycobacterium
tuberculosis. In some embodiments, the infectious disease is
tuberculosis. In certain embodiments, the bacterial infection is a
mycobacterial infection. In some embodiments the bacterial
infection is an atypical mycobacterial infection. In some
embodiments, the infectious disease is tuberculosis. In some
embodiments, the infectious disease is multi-drug resistant
tuberculosis (MDR-TB). In some embodiments, the infectious disease
is extensively drug-resistant tuberculosis (XDR-TB). In some
embodiments, the bacterial infection is caused by a member of
Staphylococcaceae. In certain embodiments, the bacterial infection
is a Staphylococcus infection. In some embodiments, the bacterial
infection is a Staphylococcus aureus infection. In some
embodiments, the bacterial infection is a methicillin-resistant
Staphylococcus aureus (MRSA) infection. In some embodiments, the
bacterial infection is healthcare-associated MRSA (HA-MRSA). In
some embodiments, the bacterial infection is community-associated
MRSA (CA-MRSA). In some embodiments, the bacterial infection is a
vancomycin-intermediate Staphylococcus aureus (VISA) infection or a
vancomycin-resistant Staphylococcus aureus (VRSA) infection. In
some embodiments, the bacterial infection is B. anthracis. In
certain embodiments, the bacterial infection is E. coli.
[0275] Exemplary bacterial infections include, but are not limited
to, infections with a Gram positive bacteria (e.g., of the phylum
Actinobacteria, phylum Firmicutes, or phylum Tenericutes); Gram
negative bacteria (e.g., of the phylum Aquificae, phylum
Deinococcus-Thermus, phylum Fibrobacteres/Chlorobi/Bacteroidetes
(FCB), phylum Fusobacteria, phylum Gemmatimonadest, phylum
Ntrospirae, phylum Planctomycetes/Verrucomicrobia/Chlamydiae (PVC),
phylum Proteobacteria, phylum Spirochaetes, or phylum
Synergistetes); or other bacteria (e.g., of the phylum
Acidobacteria, phylum Chlroflexi, phylum Chrystiogenetes, phylum
Cyanobacteria, phylum Deferrubacteres, phylum Dictyoglomi, phylum
Thermodesulfobacteria, or phylum Thermotogae).
[0276] In certain embodiments, the bacteria is a member of the
phylum Firmicutes and the genus Enterococcus, i.e., the bacterial
infection is an Enterococcus infection. Exemplary Enterococci
bacteria include, but are not limited to, E. avium, E. durans, E.
faecalis, E. faecium, E. gallinarum, E. solitarius, E.
casseliflavus, and E. raffinosus. In certain embodiments, the
bacteria is a member of the phylum Firmicutes and the genus
Staphylococcus, i.e., the bacterial infection is a Staphylococcus
infection. Exemplary Staphylococci bacteria include, but are not
limited to, S. arlettae, S. aureus, S. auricularis, S. capitis, S.
caprae, S. carnous, S. chromogenes, S. cohii, S. condimenti, S.
croceolyticus, S. delphini, S. devriesei, S. epidermis, S. equorum,
S. felis, S. fluroettii, S. gallinarum, S. haemolyticus, S.
hominis, S. hyicus, S. intermedius, S. kloosii, S. leei, S. lenus,
S. lugdunesis, S. lutrae, S. lyticans, S. massiliensis, S. microti,
S. muscae, S. nepalensis, S. pasteuri, S. penttenkoferi, S.
piscifermentans, S. psuedointermedius, S. psudolugdensis, S.
pulvereri, S. rostri, S. saccharolyticus, S. saprophyticus, S.
schleiferi, S. sciuri, S. simiae, S. simulans, S. stepanovicii, S.
succinus, S. vitulinus, S. warneri, and S. xylosus. In certain
embodiments, the Staphylococcus infection is a S. aureus infection.
In certain embodiments, the Staphylococcus infection is a
methicillin-resistant Staphylococcus aureus (MRSA) infection. In
some embodiments, the Staphylococcus infection is an
vancomycin-intermediate Staphylococcus aureus (VISA) infection or a
vancomycin-resistant Staphylococcus aureus (VRSA) infection.
[0277] In certain embodiments, the bacteria is a member of the
phylum Firmicutes and the genus Bacillus, i.e., the bacterial
infection is a Bacillus infection. Exemplary Bacillus bacteria
include, but are not limited to, B. alcalophilus, B. alvei, B.
aminovorans, B. amyloliquefaciens, B. aneurinolyticus, B.
anthracis, B. aquaemaris, B. atrophaeus, B. boroniphilus, B.
brevis, B. caldolyticus, B. centrosporus, B. cereus, B. circulans,
B. coagulans, B. firmus, B. flavothermus, B. fusiformis, B.
globigii, B. infernus, B. larvae, B. laterosporus, B. lentus, B.
licheniformis, B. megaterium, B. mesentericus, B. mucilaginosus, B.
mycoides, B. natto, B. pantothenticus, B. polymyxa, B.
pseudoanthracis, B. pumilus, B. schlegelii, B. sphaericus, B.
sporothermodurans, B. stearothermophilus, B. subtilis, B.
thermoglucosidasius, B. thuringiensis, B. vulgatis, and B.
weihenstephanensis. In certain embodiments, the Bacillus infection
is a B. subtilis infection. In certain embodiments, the B. subtilis
has an efflux (e.g., mef, msr) genotype. In certain embodiments,
the B. subtilis has a methylase (e.g., erm) genotype. In certain
embodiments, the Bacillus infection is a B. anthracis infection. In
certain embodiments, the bacteria is a member of the phylum
Firmicutes and the genus Streptococcus, i.e., the bacterial
infection is a Streptococcus infection. Exemplary Streptococcus
bacteria include, but are not limited to, S. agalactiae, S.
anginosus, S. bovis, S. canis, S. constellatus, S. dysgalactiae, S.
equinus, S. iniae, S. intermedius, S. mitis, S. mutans, S. oralis,
S. parasanguinis, S. peroris, S. pneumoniae, S. pyogenes, S. ratti,
S. salivarius, S. thermophilus, S. sanguinis, S. sobrinus, S. suis,
S. uberis, S. vestibularis, S. viridans, and S. zooepidemicus. In
certain embodiments, the Strepococcus infection is an S. pyogenes
infection. In certain embodiments, the Strepococcus infection is an
S. pneumoniae infection. In certain embodiments, the S. pneumoniae
has an efflux (e.g., mef, msr) genotype. In certain embodiments,
the S. pneumoniae has a methylase (e.g., erm) genotype. In certain
embodiments, the bacteria is a member of the phylum Firmicutes and
the genus Clostridium, i.e., the bacterial infection is a
Clostridium infection. Exemplary Clostridia bacteria include, but
are not limited to, C. botulinum, C. difficile, C. perfringens, C.
tetani, and C. sordellii.
[0278] In certain embodiments, the Gram negative bacteria is a
bacteria of the phylum Proteobacteria and the genus Escherichia.
i.e., the bacterial infection is an Escherichia infection.
Exemplary Escherichia bacteria include, but are not limited to, E.
albertii, E. blattae, E. coli, E. fergusonii, E. hermannii, and E.
vulneris. In certain embodiments, the Escherichia infection is an
E. coli infection. In certain embodiments, the Gram negative
bacteria is a bacteria of the phylum Proteobacteria and the genus
Haemophilus. i.e., the bacterial infection is an Haemophilus
infection. Exemplary Haemophilus bacteria include, but are not
limited to, H. aegyptius, H. aphrophilus, H. avium, H. ducreyi, H.
felis, H. haemolyticus, H. influenzae, H. parainfluenzae, H.
paracuniculus, H. parahaemolyticus, H. pittmaniae, Haemophilus
segnis, and H. somnus. In certain embodiments, the Haemophilus
infection is an H. influenzae infection.
[0279] In certain embodiments, the Gram negative-bacteria is a
bacteria of the phylum Proteobacteria and the genus Acinetobacter.
i.e., the bacterial infection is an Acinetobacter infection.
Exemplary Acinetobacter bacteria include, but are not limited to,
A. baumanii, A. haemolyticus, and A. lwoffii. In certain
embodiments, the Acinetobacter infection is an A. baumanii
infection. In certain embodiments, the Gram-negative bacteria is a
bacteria of the phylum Proteobacteria and the genus Klebsiella.
i.e., the bacterial infection is a Klebsiella infection. Exemplary
Klebsiella bacteria include, but are not limited to, K.
granulomatis, K. oxytoca, K. michiganensis, K. pneumoniae, K.
quasipneumoniae, and K. variicola. In certain embodiments, the
Klebsiella infection is a K. pneumoniae infection. In certain
embodiments, the Gram-negative bacteria is a bacteria of the phylum
Proteobacteria and the genus Pseudomonas. i.e., the bacterial
infection is a Pseudomonas infection. Exemplary Pseudomonas
bacteria include, but are not limited to, P. aeruginosa, P.
oryzihabitans, P. plecoglissicida, P. syringae, P. putida, and P.
fluoroscens. In certain embodiments, the Pseudomonas infection is a
P. aeruginosa infection. In certain embodiments, the Gram-negative
bacteria is a bacteria of the phylum Bacteroidetes and the genus
Bacteroides. i.e., the bacterial infection is a Bacteroides
infection. Exemplary Bacteroides bacteria include, but are not
limited to, B. fragilis, B. distasonis, B. ovatus, B.
thetaiotaomicron, and B. vulgatus. In certain embodiments, the
Bacteroides infection is a B. fragilis infection. In certain
embodiments, the Gram negative-bacteria is a bacteria of the phylum
Proteobacteria and the genus Yersinia. i.e., the bacterial
infection is an Yersinia infection. Exemplary Yersinia bacteria
include, but are not limited to, Y. pestis, Y. entercolitica. and
Y. pseudotuberculosis. In certain embodiments, the Acinetobacter
infection is an Y. pestis infection.
[0280] In certain embodiments, the bacterial infection is caused by
a bacteria of the phylum Actinobacteria. Exemplary bacteria of the
phylum include, but are not limited to bacteria within
Acidimicrobiaceae family, Actinomycetaceae family,
Corynebacteriaceae family, Gordoniaceae family, Mycobacteriaceae
family, Nocardiaceae family, Tsukamurellaceae family,
Williamsiaceae family, Acidothermaceae family, Frankiaceae family,
Geodermatophilaceae, Kineosporiaceae, Microsphaeraceae family,
Sporichthyaceae family, Glycomycetaceae family, Beutenbergiaceae
family, Bogoriellaceae family, Brevibacteriaceae family,
Cellulomonadaceae family, Dermabacteraceae family, Dermatophilaceae
family, Dermacoccaceae family, Intrasporangiaceae family,
Jonesiaceae family, Microbacteriaceae family, Micrococcaceae
family, Promicromonosporaceae family, Rarobacteraceae family,
Sanguibacteraceae family, Micromonosporaceae family,
Nocardioidaceae family, Propionibacteriaceae family,
Actinosynnemataceae family, Pseudonocardiaceae family,
Streptomycetaceae family, Nocardiopsaceae family,
Streptosporangiaceae family, Thermomonosporaceae family,
Bifidobacteriaceae family, Coriobacteriaceae family,
Rubrobacteraceae family, and Sphaerobacteraceae family.
[0281] In certain embodiments, the bacteria is a member of the
phylum Actinobacteria and the Mycobacterium. In some embodiments
the bacteria is a baceteria associated with an atypical
mycobacterial infection. Exemplary bacteria from genus
Mycobacterium include, but are not limited to: M. abscessus, M.
africanum, M. avium, M. bovis, M. caprae, M. canetti, M. chelonae,
M. colombiense, M. flavescens, M. fortuitum, M. genavense, M.
gordonae, M. haemophilum, M. intracellulare, M. kansasii, M.
leprae, M. lepramatosis, M. malmoense, M. marinum, M. microti, M.
parafortuitum, M. phlei, M. pinnipedii, M. scrofulaceum, M. simiae,
M. smegmatis, M. szulgai, M. terrae, M. ulcerans, M. vaccae, and M.
xenope. In some embodiments, the bacteria is a bacteria that can
cause tuberculosis (e.g., a member of the Mycobacterium
tuberculosis complex (e.g., M. tuberculosis, M. africanum, M.
bovis, M bovis BCG, M. microti, M. canetti, M pinnipedii, M.
suricattae, M. mungi). In some embodiments, the bacteria is M.
tuberculosis. In some embodiments, the bacteria is a member of the
Mycobacterium avium complex (e.g., M. avium, M. avium, M. avium
paratuberculosis, M. avium silvaticum, M. avium hominissuis, M.
colombiense, M. indicus pranii, M. intracellulare). In some
embodiments, the bacteria is M. phlei. In some embodiments, the
bacteria is M. smegmatis. In certain embodiments, the Mycobacterium
infection is a M. tuberculosis infection. In certain embodiments,
the Mycobacterium infection is a multi-drug-resistant tuberculosis
(MDR-TB) infection or extensively drug-resistant tuberculosis
(XDR-TB) infection. In certain embodiments, the M. tuberculosis
infection is a multi-drug-resistant tuberculosis (MDR-TB) infection
or extensively drug-resistant tuberculosis (XDR-TB) infection.
[0282] In certain embodiments, the bacterial infection is a
Mycobacterium infection, a Staphylococcus infection, Pseudomonas
infection, a Bacillus infection, or an Escherichia infection. In
certain, embodiments, the bacterial infection is tuberculosis. In
some embodiments, the bacterial infection is a Mycobacterium
tuberculosis infection. In certain embodiments, the bacterial
infection is a Pseudomonas infection. In some embodiments, the
bacterial infection is Pseudomonas aeruginosa infection. In some
embodiments, the bacterial infection is Yersinia infection. In some
embodiments the bacterial infection is Yersinia pestis infection.
In some embodiments the bacterial infection is E. coli infection.
In some embodiments the bacterial infection is Bacillus anthracis
infection. In some embodiments the bacterial infection is Bacillus
anthracis infection. In some embodiments the bacterial infection is
Vibrio cholera infection. In some embodiments, the bacterial
infection is infection of multiple species of bacterium. In some
embodiments, the bacterial infection is infection of multiple
species of bacterium, one of which is P. aeruginosa. In some
embodiments, the bacterial infection is infection of multiple
species of bacterium, one of which is Mycobacterium
tuberculosis.
[0283] In another aspect, the invention provides a method of
treating cancer in a subject, the method comprising administering
an effective amount of a compound of Formula (IX):
##STR00130##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0284] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0284] ##STR00131## [0285] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0286] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0287] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0288]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0288] ##STR00132## [0289] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0290] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0291] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0292] In another aspect, the invention provides a method of
treating cancer in a subject in need thereof, the method comprising
administering an effective amount of a compound of Formula
(IX):
##STR00133##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0293] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0293] ##STR00134## [0294] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0295] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0296] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0297]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0297] ##STR00135## [0298] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0299] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0300] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0301] In another aspect, the invention provides a method of
treating cancer in a subject, the method comprising administering
an effective amount of a compound of Formula (VII):
##STR00136##
[0302] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0303] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0303] ##STR00137## [0304] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0305] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0306] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0307] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0307] ##STR00138## [0308] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0309] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0310] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0311] In another aspect, the invention provides a method of
treating cancer in a subject in need thereof, the method comprising
administering an effective amount of a compound of Formula
(VII):
##STR00139##
[0312] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0313] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0313] ##STR00140## [0314] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0315] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0316] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0317] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0317] ##STR00141## [0318] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0319] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0320] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0321] In another aspect, the cancer is cancer of the colon,
breast, bone, brain and others (e.g., osteosarcoma, neuroblastoma,
colon adenocarcinoma), chronic myelogenous leukemia (CML), acute
myeloid leukemia (AML), acute promyelocytic leukemia (APL),
comprising administering to said subject in need thereof, an
effective amount of a compound delineated herein (e.g., any of the
formulae herein), or a pharmaceutically acceptable salt thereof.
Other cancers that may be treated by the compositions and methods
of the invention include cardiac cancer (e.g., sarcoma, myxoma,
rhabdomyoma, fibroma, lipoma and teratoma); lung cancer (e.g.,
bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma,
sarcoma, lymphoma, chondromatous hamartoma, mesothelioma); various
gastrointestinal cancer (e.g., cancers of esophagus, stomach,
pancreas, small bowel, and large bowel); genitourinary tract cancer
(e.g., kidney, bladder and urethra, prostate, testis; liver cancer
(e.g., hepatoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma,
hepatocellular adenoma, hemangioma); bone cancer (e.g., osteogenic
sarcoma, fibrosarcoma, malignant fibrous histiocytoma,
chondrosarcoma, Ewing's sarcoma, malignant lymphoma, multiple
myeloma, malignant giant cell tumor chordoma, osteochronfroma,
benign chondroma, chondroblastoma, chondromyxofibroma, osteoid
osteoma and giant cell tumors); cancers of the nervous system
(e.g., of the skull, meninges, brain, and spinal cord);
gynecological cancers (e.g., uterus, cervix, ovaries, vulva,
vagina); hematologic cancer (e.g., cancers relating to blood,
Hodgkin's disease, non-Hodgkin's lymphoma); skin cancer (e.g.,
malignant melanoma, basal cell carcinoma, squamous cell carcinoma,
Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma,
dermatofibroma, keloids, psoriasis); and cancers of the adrenal
glands (e.g., neuroblastoma).
[0322] In another aspect, the invention provides a method of
increasing the effectiveness of anti-cancer therapy in a subject
currently being administered one or more anti-cancer therapies, the
method comprising administering an effective amount of a compound
of Formula (IX):
##STR00142##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0323] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0323] ##STR00143## [0324] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0325] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0326] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0327]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0327] ##STR00144## [0328] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0329] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0330] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0331] In another aspect, the invention provides a method of
increasing the effectiveness of anti-cancer therapy in a subject
currently being administered one or more anti-cancer therapies, the
method comprising administering an effective amount of a compound
of Formula (VII):
##STR00145##
[0332] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0333] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0334] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0334] ##STR00146## [0335] R.sub.15 is H or C.sub.1-C.sub.6 alkyl;
[0336] R.sub.6 is H, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, or C.sub.2-C.sub.6 alkynyl; [0337] R.sub.8 is H, Boc,
acetyl, Fmoc, or
[0337] ##STR00147## [0338] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0339] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0340] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the subject is a mammal.
In another aspect, the subject is a human.
[0341] In another aspect, the anti-cancer therapy is an anti-cancer
agent, chemotherapeutic agent, an anti-angiogenesis agent,
cytotoxic agent, or an anti-proliferation agent. Examples of such
agents include but are not limited to TRAIL, TRAIL receptor
agonists, daunorubicin, daunomycin, dactinomycin, doxorubicin,
epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide,
ifosfamide, cytosine arabinoside, bis-chloroethylnitrosurea,
busulfan, mitomycin C, actinomycin D, mithramycin, prednisone,
hydroxyprogesterone, testosterone, tamoxifen, dacarbazine,
procarbazine, hexamethylmelamine, pentamethylmelamine,
mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea,
nitrogen mustards, melphalan, cyclophosphamide, 6-mercaptopurine,
6-thioguanine, cytarabine (CA), 5-azacytidine, hydroxyurea,
deoxycoformycin, 4-hydroxyperoxycyclophosphoramide, 5-fluorouracil
(5-FU), 5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX),
colchicine, vincristine, vinblastine, etoposide, trimetrexate,
teniposide, cisplatin and diethylstilbestrol (DES). See, generally,
The Merck Manual of Diagnosis and Therapy, 15th Ed., pp. 1206-1228,
Berkow et al., eds., Rahway, N.J., 1987). In certain embodiments,
the anti-cancer therapy is TRAIL or a TRAIL receptor agonist. In
certain embodiments, the anti-cancer therapy is TRAIL. In certain
embodiments, the anti-cancer therapy is a TRAIL receptor
agonist.
[0342] In another aspect, the invention provides a method of
increasing the effectiveness of anti-bacterial therapy in a subject
currently being administered one or more anti-bacterial therapies,
the method comprising administering an effective amount of a
compound of Formula (IX):
##STR00148##
or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug
thereof; [0343] wherein R.sub.13 is H, Boc, acetyl, Fmoc,
[0343] ##STR00149## [0344] each R.sub.14 is independently H or
C.sub.1-C.sub.6 alkyl; [0345] R.sub.15 is H or C.sub.1-C.sub.6
alkyl; [0346] each R.sub.6 is independently H, C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0347]
R.sub.8 is H, Boc, acetyl, Fmoc, or
[0347] ##STR00150## [0348] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0349] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0350] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the disease is cancer. In
another aspect, the subject is a mammal. In another aspect, the
subject is a human.
[0351] In another aspect, the invention provides a method of
increasing the effectiveness of anti-bacterial therapy in a subject
currently being administered one or more anti-bacterial therapies,
the method comprising administering an effective amount of a
compound of Formula (VII):
##STR00151##
[0352] or a pharmaceutically acceptable salt, solvate, hydrate, or
prodrug thereof; [0353] wherein R.sub.13 is H, Boc, acetyl,
Fmoc,
[0353] ##STR00152## [0354] R.sub.14 is H or C.sub.1-C.sub.6 alkyl;
[0355] R.sub.15 is H or C.sub.1-C.sub.6 alkyl; [0356] R.sub.6 is H,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6
alkynyl; [0357] R.sub.8 is H, Boc, acetyl, Fmoc, or
[0357] ##STR00153## [0358] R.sub.7 is H, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0359] R.sub.9
is H or C.sub.1-C.sub.6 alkyl; and [0360] R.sub.10 is H or
C.sub.1-C.sub.6 alkyl. In another aspect, the disease is cancer. In
another aspect, the subject is a mammal. In another aspect, the
subject is a human.
[0361] Exemplary anti-bacterial therapies include but are not
limited to gentamicin, amikacin, tobramycin, ciprofloxacin,
levofloxacin, ceftazidimine, cefepime, cefoperazone, cefpirome,
ceftobiprole, carbenicllin, ticarcillin, mezlocillin, azlocillin,
piperacillin, meropenem, imipenem, doripenem, polymyxin B,
colistin, aztreonam, isoniazid, rifampicin (also called rifampin),
pyrazinamide, ethambutol, streptomycin, moxifloxacin, gatifloxacin,
amikacin, capremycin, kanamycin, ethionamide, prothionamide,
cycloserine, terizidone, linezolide, clofazimine, pretomanid,
bedaquiline, delamanid, or rifamycins. In certain embodiments, the
additional pharmaceutical agent is isoniazid, rifampicin (also
called rifampin), pyrazinamide, ethambutol, or streptomycin. In
some embodiments, the additional pharmaceutical agent is
levofloxacin, moxifloxacin, gatifloxacin, amikacin, capremycin,
kanamycin, ethionamide, prothionamide, cycloserine, terizidone,
linezolide, or clofazimine.
[0362] In certain embodiments, the subject is a mammal, preferably
a primate or human.
[0363] Methods delineated herein include those wherein the subject
is identified as in need of a particular stated treatment.
Identifying a subject in need of such treatment can be in the
judgment of a subject or a health care professional and can be
subjective (e.g. opinion) or objective (e.g. measurable by a test
or diagnostic method).
[0364] In another embodiment, the invention provides a method as
described above, wherein the effective amount of the compound of
any of the formulae herein ranges from about 0.005 .mu.g/kg to
about 200 mg/kg. In certain embodiments, the effective amount of
the compound of any of the formulae herein ranges from about 0.1
mg/kg to about 200 mg/kg. In a further embodiment, the effective
amount of compound of any of the formulae herein ranges from about
10 mg/kg to 100 mg/kg.
[0365] In other embodiments, the invention provides a method as
described above wherein the effective amount of the compound of any
of the formulae herein ranges from about 1.0 pM to about 500 nM. In
certain embodiments, the effective amount ranges from about 10.0 pM
to about 1000 pM. In another embodiment, the effective amount
ranges from about 1.0 nM to about 10 nM.
[0366] In another embodiment, the invention provides a method as
described above, wherein the compound of any of the formulae herein
is administered intravenously, intramuscularly, subcutaneously,
intracerebroventricularly, orally or topically.
[0367] In other embodiments, the invention provides a method as
described above, wherein the compound of any of the formulae herein
is administered alone or in combination with one or more other
therapeutics. In a further embodiment, the additional therapeutic
agent is an anti-bacterial agent. Examples of such anti-bacterial
agents include but are not limited to gentamicin, amikacin,
tobramycin, ciprofloxacin, levofloxacin, ceftazidimine, cefepime,
cefoperazone, cefpirome, ceftobiprole, carbenicllin, ticarcillin,
mezlocillin, azlocillin, piperacillin, meropenem, imipenem,
doripenem, polymyxin B, colistin, aztreonam, isoniazid, rifampicin
(also called rifampin), pyrazinamide, ethambutol, streptomycin,
moxifloxacin, gatifloxacin, amikacin, capremycin, kanamycin,
ethionamide, prothionamide, cycloserine, terizidone, linezolide,
clofazimine, pretomanid, bedaquiline, delamanid, or rifamycins. In
certain embodiments, the additional pharmaceutical agent is
isoniazid, rifampicin (also called rifampin), pyrazinamide,
ethambutol, or streptomycin. In some embodiments, the additional
pharmaceutical agent is levofloxacin, moxifloxacin, gatifloxacin,
amikacin, capremycin, kanamycin, ethionamide, prothionamide,
cycloserine, terizidone, linezolide, or clofazimine.
[0368] In other embodiments, the invention provides a method as
described above, wherein the compound of any of the formulae herein
is administered alone or in combination with one or more other
therapeutics. In a further embodiment, the additional therapeutic
agent is an anti-cancer agent, chemotherapeutic agent, an
anti-angiogenesis agent, cytotoxic agent, or an anti-proliferation
agent. Examples of such chemotherapeutic agents include but are not
limited to daunorubicin, daunomycin, dactinomycin, doxorubicin,
epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide,
ifosfamide, cytosine arabinoside, bis-chloroethylnitrosurea,
busulfan, mitomycin C, actinomycin D, mithramycin, prednisone,
hydroxyprogesterone, testosterone, tamoxifen, dacarbazine,
procarbazine, hexamethylmelamine, pentamethylmelamine,
mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea,
nitrogen mustards, melphalan, cyclophosphamide, 6-mercaptopurine,
6-thioguanine, cytarabine (CA), 5-azacytidine, hydroxyurea,
deoxycoformycin, 4-hydroxyperoxycyclophosphoramide, 5-fluorouracil
(5-FU), 5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX),
colchicine, vincristine, vinblastine, etoposide, trimetrexate,
teniposide, cisplatin and diethylstilbestrol (DES). See, generally,
The Merck Manual of Diagnosis and Therapy, 15th Ed., pp. 1206-1228,
Berkow et al., eds., Rahway, N.J., 1987).
[0369] Another object of the present invention is the use of a
compound as described herein (e.g., of any formulae herein) in the
manufacture of a medicament for use in the treatment of a disorder
or disease (e.g., cancer or bacterial infection). Another object of
the present invention is the use of a compound as described herein
(e.g., of any formulae herein) for use in the treatment of a
disorder or disease (e.g., cancer or bacterial infection).
Pharmaceutical Compositions
[0370] In one aspect, the invention provides a pharmaceutical
composition comprising the compound of any of the formulae herein
(e.g., Formulae (I)-(IX)) or pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier. In another
aspect, the composition further comprises an additional agent. In
another aspect, the additional agent is an anti-bacterial agent. In
another aspect, the additional agent is an anti-cancer agent.
[0371] In one aspect, the invention provides a kit comprising an
effective amount of a compound of any of the formulae herein (e.g.,
Formulae (I)-(IX)), in unit dosage form, together with instructions
for administering the compound to a subject suffering from or
susceptible to a bacterial infection.
[0372] In one aspect, the invention provides a kit comprising an
effective amount of a compound of any of the formulae herein (e.g.,
Formulae (I)-(IX)), in unit dosage form, together with instructions
for administering the compound to a subject suffering from or
susceptible to cancer.
[0373] The term "pharmaceutically acceptable salts" or
"pharmaceutically acceptable carrier" is meant to include salts of
the active compounds which are prepared with relatively nontoxic
acids or bases, depending on the particular substituents found on
the compounds described herein. When compounds of the present
invention contain relatively acidic functionalities, base addition
salts can be obtained by contacting the neutral form of such
compounds with a sufficient amount of the desired base, either neat
or in a suitable inert solvent. Examples of pharmaceutically
acceptable base addition salts include sodium, potassium, calcium,
ammonium, organic amino, or magnesium salt, or a similar salt. When
compounds of the present invention contain relatively basic
functionalities, acid addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired acid, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable acid addition salts include those
derived from inorganic acids like hydrochloric, hydrobromic,
nitric, carbonic, monohydrogencarbonic, phosphoric,
monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, maleic, malonic, benzoic,
succinic, suberic, fumaric, lactic, mandelic, phthalic,
benzenesulfonic, p-tolylsulfonic, citric, tartaric,
methanesulfonic, and the like. Also included are salts of amino
acids such as arginate and the like, and salts of organic acids
like glucuronic or galactunoric acids and the like (see, e.g.,
Berge et al., Journal of Pharmaceutical Science 66:1-19 (1977)).
Certain specific compounds of the present invention contain both
basic and acidic functionalities that allow the compounds to be
converted into either base or acid addition salts. Other
pharmaceutically acceptable carriers known to those of skill in the
art are suitable for the present invention.
[0374] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents, but otherwise the
salts are equivalent to the parent form of the compound for the
purposes of the present invention.
[0375] In addition to salt forms, the present invention provides
compounds which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0376] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present invention and are intended to be within the scope of the
present invention.
[0377] The invention also provides a pharmaceutical composition,
comprising an effective amount a compound described herein and a
pharmaceutically acceptable carrier. In an embodiment, compound is
administered to the subject using a pharmaceutically-acceptable
formulation, e.g., a pharmaceutically-acceptable formulation that
provides sustained delivery of the compound to a subject for at
least 12 hours, 24 hours, 36 hours, 48 hours, one week, two weeks,
three weeks, or four weeks after the pharmaceutically-acceptable
formulation is administered to the subject.
[0378] Actual dosage levels and time course of administration of
the active ingredients in the pharmaceutical compositions of this
invention may be varied so as to obtain an amount of the active
ingredient which is effective to achieve the desired therapeutic
response for a particular patient, composition, and mode of
administration, without being toxic (or unacceptably toxic) to the
patient.
[0379] In use, at least one compound according to the present
invention is administered in a pharmaceutically effective amount to
a subject in need thereof in a pharmaceutical carrier by
intravenous, intramuscular, subcutaneous, or intracerebro
ventricular injection or by oral administration or topical
application. In accordance with the present invention, a compound
of the invention may be administered alone or in conjunction with a
second, different therapeutic. By "in conjunction with" is meant
together, substantially simultaneously or sequentially. In one
embodiment, a compound of the invention is administered acutely.
The compound of the invention may therefore be administered for a
short course of treatment, such as for about 1 day to about 1 week.
In another embodiment, the compound of the invention may be
administered over a longer period of time to ameliorate chronic
disorders, such as, for example, for about one week to several
months depending upon the condition to be treated.
[0380] By "pharmaceutically effective amount" as used herein is
meant an amount of a compound of the invention, high enough to
significantly positively modify the condition to be treated but low
enough to avoid serious side effects (at a reasonable benefit/risk
ratio), within the scope of sound medical judgment. A
pharmaceutically effective amount of a compound of the invention
will vary with the particular goal to be achieved, the age and
physical condition of the patient being treated, the severity of
the underlying disease, the duration of treatment, the nature of
concurrent therapy and the specific apratoxin compound employed.
For example, a therapeutically effective amount of a compound of
the invention administered to a child or a neonate will be reduced
proportionately in accordance with sound medical judgment. The
effective amount of a compound of the invention will thus be the
minimum amount which will provide the desired effect.
[0381] The compound may be administered parenterally or
intraperitoneally. Dispersions can also be prepared, for example,
in glycerol, liquid polyethylene glycols, and mixtures thereof, and
in oils.
[0382] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions (where water soluble) or dispersions and
sterile powders for the extemporaneous preparation of sterile
injectable solutions or dispersions. In all cases the form must be
sterile and must be fluid to the extent that easy syringability
exists. It must be stable under the conditions of manufacture and
storage. The carrier can be a solvent or dispersion medium
containing, for example, water, DMSO, ethanol, polyol (for example,
glycerol, propylene glycol, liquid polyethylene glycol, and the
like), suitable mixtures thereof and vegetable oils. The proper
fluidity can be maintained, for example, by the use of a coating
such as lecithin, by the maintenance of the required particle size
in the case of dispersion. In many cases it will be preferable to
include isotonic agents, for example, sugars or sodium chloride.
Prolonged absorption of the injectable compositions can be brought
about by the use in the compositions of agents delaying absorption,
for example, aluminum monostearate and gelatin.
[0383] Sterile injectable solutions are prepared by incorporating
the compound of the invention in the required amount in the
appropriate solvent with various of the other ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the various
sterilized compounds into a sterile vehicle which contains the
basic dispersion medium and the required other ingredients from
those enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, the preferred methods
of preparation are vacuum-drying and the freeze-drying technique
which yields a powder of the active ingredient plus any additional
desired ingredient from previously sterile-filtered solution
thereof.
[0384] For oral therapeutic administration, the compound may be
incorporated with excipients and used in the form of ingestible
tablets, buccal tablets, troches, capsules, elixirs, suspensions,
syrups, wafers, and the like. Compositions or preparations
according to the present invention are prepared so that an oral
dosage unit form contains compound concentration sufficient to
treat a disorder in a subject.
[0385] Some examples of substances which can serve as
pharmaceutical carriers are sugars, such as lactose, glucose and
sucrose; starches such as corn starch and potato starch; cellulose
and its derivatives such as sodium carboxymethycellulose,
ethylcellulose and cellulose acetates; powdered tragancanth; malt;
gelatin; talc; stearic acids; magnesium stearate; calcium sulfate;
vegetable oils, such as peanut oils, cotton seed oil, sesame oil,
olive oil, corn oil and oil of theobroma; polyols such as propylene
glycol, glycerine, sorbitol, manitol, and polyethylene glycol;
agar; alginic acids; pyrogen-free water; isotonic saline; and
phosphate buffer solution; skim milk powder; as well as other
non-toxic compatible substances used in pharmaceutical formulations
such as Vitamin C, estrogen and echinacea, for example. Wetting
agents and lubricants such as sodium lauryl sulfate, as well as
coloring agents, flavoring agents, lubricants, excipients,
tableting agents, stabilizers, anti-oxidants and preservatives, can
also be present.
[0386] The recitation of a listing of chemical groups in any
definition of a variable herein includes definitions of that
variable as any single group or combination of listed groups. The
recitation of an embodiment for a variable herein includes that
embodiment as any single embodiment or in combination with any
other embodiments or portions thereof. The recitation of an
embodiment herein includes that embodiment as any single embodiment
or in combination with any other embodiments or portions
thereof.
Examples
[0387] The present invention will now be demonstrated using
specific examples that are not to be construed as limiting.
General Experimental Procedures
[0388] All commercial reagents were used without further
purification unless otherwise noted. Solvents were purified
according to the guidelines in Purification of Laboratory Chemicals
(5.sup.th edition, W. L. F. Armarego, Christina L. L. Chai,
Butterworth-Heinemann: Oxford, 2003). Tetrahydrofuran (THF),
CH.sub.2Cl.sub.2, DMF were purified by MS-PDS solvent purification
system (Innovation Inc.). All reactions were performed in heat-gun
dried flasks (400.degree. C. under reduced pressure) under an inert
atmosphere of anhydrous Ar unless otherwise noted. Thin layer
chromatography was performed on EMD silica gel 60 .ANG. F.sub.254
glass plates and preparative thin layer chromatography was
performed on Whatman silica gel 60 .ANG. F.sub.254 glass plates
(layer thick 1000 .mu.m). Flash column chromatography was performed
with Fisher 170-400 mesh silica gel. Nuclear magnetic resonance
(NMR) spectra were recorded on a Bruker Avance III 500
spectrometer, a Bruker Avance III 600 MHz spectrometer or a Bruker
Avance Neo-600 spectrometer with a broadband Prodigy cryogenic
probe. Chemical shifts for proton nuclear magnetic resonance
(.sup.1H NMR) spectra are reported in parts per million relative to
the signal residual CDCl.sub.3 at 7.26 ppm; Chemicals shifts for
carbon nuclear magnetic resonance (.sup.13C NMR) spectra are
reported in parts per million relative to the center line of the
CDCl.sub.3 triplet at 77.16 ppm; The abbreviations s, d, dd, ddd,
dddd, t, q, p, br, and m stand for the resonance multiplicity
singlet, doublet, doublet of doublets, doublet of doublet of
doublets, doublet of doublet of doublet of doublets, triplet,
quartet, pentet, broad and multiplet, respectively. Optical
rotation was measured on a Perkin-Elmer 341 polarimeter (Na D line)
using a microcell of 1 dm path length. HRMS was conducted using a
Thermo Fisher Q Exactive Focus mass spectrometer equipped with
UltiMate.TM. 3000 RSLCnano System and electrospray probe on
Universal Ion Max API source. Fluorescence and UV were measured on
a SpectraMax M5 (Molecular Devices).
Isolation of Doscadenamide A
[0389] A cyanobacterium Moorea bouillonii sample was collected at
Finger's Reef, Guam and previous investigation of this sample has
led to the isolation of apratoxin A, lyngbyaloside,
2-epi-lyngbyaloside, 18E-lyngbyaloside C, 18Z-lyngbyaloside C, and
apratyramide [Luesch, H.; Yoshida, W. Y.; Moore, R. E.; Paul, V.
J.; Corbett, T. H. J. Am. Chem. Soc. 2001, 123, 5418-5423; Matthew,
S.; Salvador, L. A.; Schupp, P. J.; Paul, V. J.; Luesch, H., J.
Nat. Prod. 2010, 73 (9), 1544-1552; Cal, W.; Salvador-Reyes, L. A.;
Zhang, W.; Chen, Q. Y.; Matthew, S.; Ratnayake, R.; Seo, S. J.;
Dolles, S.; Gibson, D. J.; Paul, V. J.; Luesch, H., ACS Chem. Biol.
2018, 13 (1), 91-99]. The cyanobacterial sample was fractionated as
described previously and the isolation was achieved by silica gel
column chromatography and several rounds of reversed-phase HPLC to
yield doscadenamide A (1a), as illustrated in FIG. 1 {white solid,
[.alpha.].sup.20.sub.D 40 (c 0.07, MeOH)} [Matthew, S.; Salvador,
L. A.; Schupp, P. J.; Paul, V. J.; Luesch, H., J. Nat. Prod. 2010,
73 (9), 1544-1552]. Specifically, the sample of the Moorea
bouillonii cyanobacterium was extracted with CH.sub.2Cl.sub.2 and
MeOH (2:1) and the extract (10 g) was fractionated using column
chromatography on silica gel, eluting with CH.sub.2Cl.sub.2
containing increasing concentrations of iPrOH to afford 16
fractions. Fractions 3, 4 and 5 (2% iPrOH in CH.sub.2Cl.sub.2; 400
mg, 1.54 g and 100 mg) and fractions 6 (5% iPrOH in
CH.sub.2Cl.sub.2; 71 mg) were individually subjected to
semipreparative HPLC (Phenomenex Phenyl-hexyl, 250.times.10 mm,
5.mu., 2.0 mL/min; PDA detection) using a MeOH--H.sub.2O linear
gradient (90-100% MeOH in 30 min and 100% MeOH for 10 min).
Fractions were pooled based on retention times, .sup.1H NMR
analysis, and low-resolution MS measurements to afford impure
doscadenamide A (1a)-containing fractions. These fractions were
further purified with HPLC (Ultracarb, 250.times.10 mm, 5.mu., 2.0
mL/min; PDA detection) using a MeOH--H.sub.2O linear gradient
(90-100% MeOH in 30 min and 100% MeOH for 10 min) to afford
doscadenamide A (1a, t.sub.R=10.0 min, 4.5 mg).
Structural Elucidation of Doscadenamide A
[0390] The HRESIMS of 1a in the positive mode exhibited a
[M+H].sup.+ peak at m/z 457.3066, which suggested a molecular
formula C.sub.27H.sub.40N.sub.2O.sub.4 with 9 degrees of
unsaturation. The structure of 1a was elucidated using a
combination of 1D and 2D NMR techniques. The .sup.1H and .sup.13C
NMR spectra in CDCl.sub.3 (FIGS. 2 and 3) indicated the presence of
several characteristic signals corresponding to one O-methyl group
(.delta.H 3.85 ppm, .delta.C 58.9 ppm), two alkyne groups (.delta.H
1.92-1.94 ppm, .delta.C 68.5, 68.5, 84.6, 84.7 ppm), one
.alpha.-proton (.delta.H 4.64-4.66 ppm, .delta.C 59.2 ppm), two
.alpha.-methyl groups (.delta.H 1.12-1.14 ppm, .delta.C 16.3-18.1
ppm) and several methylene groups (.delta.H 1.30-1.90, 2.06-2.18
ppm, .delta.C 18.4-39.4 ppm). Examination of the 2D NMR spectra
(COSY, TOCSY, HSQC, HMBC and NOESY, FIGS. 4-8, and Table 1) in
CDCl.sub.3 revealed the structural skeleton of 1a.
TABLE-US-00001 TABLE 1 .sup.1H and .sup.13C NMR spectral data of
doscadenamide A (1a) at 600 MHz and 150 MHz in CDCl.sub.3 at
27.degree. C. (.delta. in ppm, J in Hz). C/H Unit No. .delta.H (J)
.delta.C COSY HMBC.sup.a pyLys- 1 170.0 OMe 2 5.05, s 94.2 1, 9 3
179.2 4 4.65, dd 59.2 5 2, 3, 5, 6 (5.4, 3.0) 5a 2.07, ddt 29.0 4,
5b, 6 3, 6, 7, 9 (13.8, 11.4, 5.4) 5b 1.85, dddd 4, 5a, 6 3, 6, 7,
9 (13.8, 11.4, 5.4, 3.0) 6a 1.19, m 20.4 5, 6b, 7 8, 10 6b 1.15, m
5, 6a, 7 8, 10 7 1.47, m 29.6 6, 8 5, 6, 8 8a 3.24, m 39.4 NH, 7 8b
3.15, m NH, 7 5, 6, 10 9 3.85, s 58.9 3 NH 5.47, br s 8 8, 10 Moya1
10 177.0 11 2.13, m 41.8 12, 18 10, 12, 18 12a 1.63, m 33.9 12b, 11
10, 11, 14, 18 12b 1.38, m 12a, 11 10, 11, 14, 18 13a 1.43, m 28.5
12, 13b, 14 12, 14 13b 1.40, m 12, 13a, 14 12, 14 14a 1.51, m 28.5
13, 14b, 15 12, 15, 17 14b 1.37, m 13, 14a, 15 12, 15, 17 15 2.16,
m 18.4 14, 17 14, 16, 17 16 84.6 17 1.93, t (2.6) 68.5 15 14 18
1.13, d (6.6) 18.1 11 10, 11 Moya2 19 176.4 20 3.77, sextet (6.6)
39.1 21, 27 19, 21, 27 21a 1.75, m 33.7 20, 21b, 22 20, 22, 27 21b
1.42, m 20, 21a, 22 20, 22, 27 22a 1.51, m 26.7 21, 22b, 23 21, 23,
24, 25 22b 1.37, m 21, 22a, 23 21, 23, 24, 25 23a 1.51 26.4 22,
23b, 24 22, 24, 25 23b 1.49 22, 23a, 24 22, 24, 25 24 2.17, m 18.5
23, 26 22, 23, 25, 26 25 84.7 26 1.93, t (2.6) 68.5 24 24 27 1.12,
d (6.6) 16.3 20 19, 20, 21 .sup.aHMBC correlations are from proton
stated to the indicated carbons.
[0391] To establish the absolute configuration, Doscadenamide A 1a
(2 mg) was dissolved in 3 mL of CH.sub.2Cl.sub.2 and ozone was
bubbled through the solution for 30 min at 25.degree. C. The
solvent was then evaporated and the residue was suspended in
H.sub.2O.sub.2--HCOOH (1:2) and heated for 20 min at 70.degree. C.
(Scheme 1).
##STR00154##
[0392] Subsequently, the resulting mixture was concentrated to
dryness and subjected to acid hydrolysis with 6N HCl. The
hydrolysate was concentrated and partitioned between EtOAc and
water. The aqueous layer was then subjected to chiral HPLC analysis
(Phenomenex, Chirex 3126 N,S-dioctyl-(D)-penicillamine, 250
mm.times.4.60 mm, 5 .mu.m; 1 mM CuSO.sub.4 in MeCN; 1.0 mL/min;
detection by UV at 254 nm). The absolute configuration of the
lysine unit was established as L-Lys (6.5 min), while the authentic
D-Lys standard eluted at 8.5 min. The EtOAc layer was coupled with
R- or S-phenylglycine methyl ester (PGME) (Scheme 2) to afford the
S-PGME derivative {HRMS (ESI) m/z calcd for
C.sub.26H.sub.33NO.sub.3 [M+H].sup.+ 469.2333, found 469.2331} or
R-PGME derivative {HRMS (ESI) m/z calcd for
C.sub.26H.sub.33NO.sub.3 [M+H].sup.+ 469.2333, found 469.2332} of
the resulting .alpha.-chiral carboxylic acid, respectively.
##STR00155##
[0393] The .DELTA..delta. values of the methyl protons (+0.05,
.DELTA..delta.=.delta..sub.S-.delta..sub.R, FIG. 9), the
.alpha.-methine proton (-- 0.03,
.DELTA..delta.=.delta..sub.S-.delta..sub.R, FIG. 9) and the
adjacent methylene protons (-0.02, -0.06,
.DELTA..delta.=.delta..sub.S-.delta..sub.R, FIG. 9) in the
resulting carboxylic acid indicated the configuration of the
.alpha.-methine in the side chain of 1a is highly likely R--.
However, during the investigation, a minor diastereomer signal was
found in addition to the major NMR signal corresponding to the
.alpha.-methyl group, which could be observed evidently from the
COSY spectra of the two PGME derivatives (Figures S8 and S9). To
further validate the configuration of 1a and provide sufficient
material for thorough biological investigation, the total synthesis
of 1a was accomplished.
Total Synthesis of Doscadenamide A
[0394] As depicted in Scheme 3, the retrosynthetic analysis of 1a
relied on the disconnection at the two amide linkage between the
pyrrolinone ring and two side chain carboxylic acids, which in the
case of 1a are the same as (R)-2-methyloct-7-ynoic acid (Moya, 2a).
The pyrrolinone ring can be obtained via the reaction between the
double protected amino acid Fmoc-L-Lys (Boc)-OH (5) and Meldrum's
acid (6).
##STR00156##
[0395] As for the synthesis of 2a, the target compound can be
achieved in 14 steps using a method reported in 2005 [Chen, H.;
Feng, Y.; Xu, Z.; Ye, T., Tetrahedron 2005, 61 (47), 11132-11140].
To improve the efficiency and introduce more flexibility into the
production of diverse carboxylic acids with .alpha.-substituted
alkyl groups, an optimized synthetic method was developed (Scheme
4), where 2a can be obtained in 4 steps with 45% overall yield. The
commercially available oct-7-ynoic acid (2c) was activated using
pivaloyl chloride followed by addition of the lithium salt of the
oxazolidinone chiral auxiliary at -78.degree. C. The resulting 7a
was methylated under conventional conditions to yield 8a as single
diastereomer [Evans, D.; Ennis, M.; Mathre, D., J. Am. Chem. Soc.
1982, 104 (6), 1737-1739]. The target compound 2a was obtained
following alkaline hydrolysis by lithium hydroperoxide [Evans, D.;
Britton, T.; Ellman, J., Tetrahedron Lett. 1987, 28 (49),
6141-6144].
##STR00157##
[0396] The total synthesis of 1a was accomplished using the
synthetic carboxylic acid 2a (Scheme 5), including generation of
the pyrrolinone core and two coupling processes to assemble the
structure. The pyrrolidine-2,4-dione 9a was prepared through
condensation of 5 with 6 in the presence of EDCI and DMAP, followed
by thermolysis [Hosseini, M.; Kringelum, H.; Murray, A.; Tonder,
J., Org. Lett. 2006, 8 (10), 2103-2106]. This intermediate was used
in the next step without purification. Conversion of 9a into its
O-methylated derivative 4 was achieved by treatment with
trimethylsilyldiazomethane. Subsequently, the N-Fmoc protecting
group in 4 was removed using piperidine to yield the secondary
amide of tetramic acid 3. The first coupling was accomplished by
condensation of the anion derived from deprotonation of 3 and the
active ester 10a derived from activation of 2a by pentafluorophenol
to yield 11a [Jin, Y.; Liu, Y.; Wang, Z.; Kwong, S.; Xu, Z.; Ye,
T., Org. Lett. 2010, 12 (5), 1100-1103]. After removal of the
Boc-protecting group with TFA, the intermediate 12a was condensed
with 2a using typical coupling conditions to afford the target
compound 1a in 6 steps with 30% overall yield from 3.
##STR00158##
[0397] To further validate the configuration of the stereocenters
in the side chain, the other three diastereomers (1b, 1c and 1d,
FIG. 12) of 1a were also synthesized using the described synthetic
method. From comparison of the NMR spectra of all the four
diastereomers with those of the isolated natural product
doscadenamide A (FIGS. 9, 13 and 14), the four diastereomers
displayed different .sup.1H NMR signals in .delta. 5.42-5.55 ppm,
.delta. 3.11-3.28 ppm, .delta. 2.04-2.21 ppm, .delta. 1.90-1.95
ppm, .delta. 1.32-1.56 ppm and .delta. 1.09-1.22 ppm. Only the
.sup.1H and .sup.13C NMR spectra of the synthetic 1a matched those
of the natural product. Meanwhile, 1a and 1b exhibited virtually
the same .sup.13C NMR spectrum as well as the natural product,
while 1c and 1d displayed different .sup.13C NMR spectra from the
isolated doscadenamide A. In addition, the optical rotation values
of 1a {[.alpha.].sup.20.sub.D 54.3 (c 0.07, MeOH)}, 1b
{[.alpha.].sup.20.sub.D 62.1 (c 0.07, MeOH)}, 1c
{[.alpha.].sup.20.sub.D 95.0 (c 0.07, MeOH)} and 1d
{[.alpha.].sup.20.sub.D 60.0 (c 0.07, MeOH)} further confirmed that
the absolute configurations of synthetic diastereomers are
consistent with the isolated doscadenamide A. Therefore, the
proposed configuration of doscadenamide A was confirmed.
Preparation of Compounds 7a and 7b
##STR00159##
[0399] To a solution of 7-octynoic acid (2c) (492.6 mg, 3.51 mmol)
and trimethylamine (TEA) (0.64 mL, 4.57 mmol) in THF (20 mL) at
-20.degree. C. was added neat pivaloyl chloride (0.48 mL, 3.87
mmol) dropwise over 20 min. The resulting mixture was stirred at
-20.degree. C. for 30 min and 0.degree. C. for another 30 min, then
it was cooled to -78.degree. C. In another reaction flask,
n-butyllithium (n-BuLi) (1.6 M in n-hexane) (2.2 ml, 3.51 mmol) was
added dropwise to a solution of (R)-oxazolidione (622.6 mg, 3.51
mmol) in tetrahydrofuran at -78.degree. C. The mixture was stirred
at this temperature for 20 min and then transferred to the above
solution of 2c in THF at -78.degree. C. by cannula. The resulting
mixture was stirred at this temperature for 30 min, then it was
allowed to warm to room temperature and stirred for additional 1.5
h. The reaction was quenched with saturate NH.sub.4Cl solution,
followed by extraction with EtOAc (50 mL.times.3). The organic
layer was then washed with 5% NaHCO.sub.3 aqueous solution, dried
over anhydrous MgSO.sub.4 and concentrated in vacuo. The residue
was purified by silica gel chromatography column (eluted by 15%
ethyl acetate in hexane) to afford 7a (918.0 mg, 87%) as white
solid. Intermediate 7b was obtained following the same synthetic
procedures using auxiliary (S)-oxazolidione to construct the
desired configuration.
(R)-4-benzyl-3-(oct-7-ynoyl)oxazolidin-2-one (7a)
[0400] [.alpha.].sup.20.sub.D -96.0 (c 0.1, MeOH); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm: 7.29 (t, J=7.5 Hz, 2H), 7.23 (t,
J=7.5 Hz, 1H), 7.17 (d, J=7.5 Hz, 2H), 4.65-4.60 (m, 1H), 4.16-4.10
(m, 2H), 3.24 (dd, J=13.5, 3.0 Hz, 1H), 2.98-2.82 (m, 2H), 2.74
(dd, J=13.5, 9.5 Hz, 1H), 2.17 (td, J=7.0, 2.5 Hz, 2H), 1.92 (t,
J=2.5 Hz, 1H), 1.71-1.64 (m, 2H), 1.55 (p, J=7.0, 2H), 1.47 (p,
J=7.5, 2H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. ppm: 173.0,
153.3, 135.3, 129.3, 128.8, 127.2, 84.3, 68.4, 66.1, 55.0, 37.8,
35.3, 28.1, 28.1, 23.6, 18.2; HRMS (ESI) m/z calcd for
C.sub.18H.sub.22NO.sub.3 [M+H].sup.+ 300.1594, found 300.1590.
(S)-4-benzyl-3-(oct-7-ynoyl)oxazolidin-2-one (7b)
[0401] (802.0 mg, 95%); [.alpha.].sup.20.sub.D 91.0 (c 0.1, MeOH);
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. ppm: 7.33 (t, J=7.2 Hz,
2H), 7.27 (t, J=7.2 Hz, 1H), 7.20 (d, J=6.6 Hz, 2H), 4.67 (ddt,
J=8.7, 8.7, 3.0 Hz, 1H), 4.21-4.15 (m, 2H), 3.29 (dd, J=13.2, 3.6
Hz, 1H), 3.01-2.88 (m, 2H), 2.77 (dd, J=13.2, 9.6 Hz, 1H), 2.21
(td, J=7.2, 2.4 Hz, 2H), 1.94 (t, J=2.4 Hz, 1H), 1.76-1.68 (m, 2H),
1.63-1.56 (m, 2H), 1.53-1.49 (m, 2H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. ppm: 173.3, 153.6, 135.4, 129.5, 129.1, 127.5,
84.5, 68.5, 66.3, 55.2, 38.0, 35.5, 28.3, 28.3, 23.8, 18.4; HRMS
(ESI) m/z calcd for C.sub.18H.sub.22NO.sub.3 [M+H].sup.+ 300.1594,
found 300.1591.
Preparation of Compounds 8a and 8b
##STR00160##
[0403] To a solution of sodium bis(trimethylsilyl)amide (NaHMDS)
(2.0 M in THF) (1.69 ml, 3.37 mmol) in anhydrous THF (10 ml) at
-78.degree. C. was added compound 7a (918.0 mg, 3.07 mmol) in THF
(5.0 ml) under argon atmosphere. After stirring at the same
temperature for 30 min, neat MeI (0.96 ml, 15.35 mmol) was added
dropwise over 10 min to the resulting reaction solution. The
reaction was then quenched with saturate NH.sub.4Cl (aq) (20 ml)
after it was stirred at -78.degree. C. for 20 h. Subsequently, the
residue was extracted with ethyl acetate (30 ml.times.3), dried
over anhydrous MgSO.sub.4, and concentrated in vacuo. The residue
was purified by silica gel chromatography column (eluted by 8%
ethyl acetate in hexane) to afford products 8a (493.8 mg, 51%) as
white solid. Intermediate 8b was obtained using 7b following the
same synthetic procedures.
(R)-4-benzyl-3-((R)-2-methyloct-7-ynoyl)oxazolidin-2-one (8a)
[0404] [.alpha.].sup.20.sub.D -268.5 (c 0.1, MeOH); .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm: 7.33 (t, J=7.5 Hz, 2H), 7.27 (t,
J=7.5 Hz, 1H), 7.21 (d, J=7.5 Hz, 2H), 4.70-4.65 (m, 1H), 4.22-4.15
(m, 2H), 3.71 (sextet, J=6.5 Hz, 1H), 3.26 (dd, J=13.5, 3.5 Hz,
1H), 2.77 (dd, J=13.5, 10.0 Hz, 1H), 2.20-2.17 (m, 2H), 1.93 (br t,
1H), 1.79-1.71 (m, 1H), 1.56-1.50 (m, 2H), 1.48-1.38 (m, 3H), 1.23
(d, J=6.5 Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. ppm:
177.3, 153.2, 135.4, 129.6, 129.1, 127.5, 84.5, 68.5, 66.2, 55.5,
38.1, 37.8, 32.9, 28.5, 26.5, 18.4, 17.5; HRMS (ESI) m/z calcd for
C.sub.19H.sub.24NO.sub.3 [M+H].sup.+ 314.1751, found 314.1748.
(S)-4-benzyl-3-((R)-2-methyloct-7-ynoyl)oxazolidin-2-one (8b)
[0405] (378.3 mg, 45%); [.alpha.].sup.20.sub.D 18.0 (c 0.1, MeOH);
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. ppm: 7.33-7.31 (m, 2H),
7.29-7.26 (m, 1H), 7.22-7.20 (m, 2H), 4.68 (ddt, J=9.6, 7.8, 3.0
MHz, 1H), 4.22-4.16 (m, 2H), 3.71 (h, J=7.2 Hz, 1H), 3.26 (dd,
J=13.2, 3.6 Hz, 1H), 2.77 (dd, J=13.2, 9.0 Hz, 1H), 2.19 (td,
J=6.6, 1.8 Hz, 2H), 1.93 (t, J=2.4 Hz, 1H), 1.80-1.72 (m, 1H),
1.58-1.50 (m, 2H), 1.49-1.39 (m, 3H), 1.23 (d, J=6.6 Hz, 3H);
.sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm: 177.2, 153.2,
135.4, 129.6, 129.1, 127.5, 84.5, 68.5, 66.2, 55.5, 38.0, 37.8,
32.9, 28.5, 26.5, 18.4, 17.5; HRMS (ESI) m/z calcd for
C.sub.19H.sub.24NO.sub.3[M+H].sup.+ 314.1751, found 314.1749.
Preparation of Compounds 2a and 2b
##STR00161##
[0407] Hydrogen peroxide (30% in H.sub.2O.sub.2) (0.55 ml, 4.82
mmol) was added to the solution of 8a (378.8 mg, 1.20 mmol) in the
mixture of THF-H.sub.2O (8.0 ml-4.0 ml) at 0.degree. C. After
stirring at 0.degree. C. for 10 min, LiOH.H.sub.2O (101.1 mg, 2.41
mmol) was added to the above reaction solution. Then the resulting
reaction mixture was stirred at 0.degree. C. for 2 h and additional
1 h at room temperature before Na.sub.2SO.sub.3 (760 mg) was added.
The quenched reaction mixture was diluted with water (20 ml) and
EtOAc (20 ml). The organic ethyl acetate phase was separated and
the aqueous phase was acidified with 1M HCl(aq) to pH 2 and
extracted EtOAc (20 ml.times.3). The combined EtOAc phase was dried
over anhydrous MgSO.sub.4, concentrated in vacuo and purified by
silica gel column chromatography (eluted by 17% ethyl acetate in
hexane) to afford products 2a (175.0 mg, 94%) as clear oil. 2b was
obtained from 8b following the same procedure.
(R)-2-methyloct-7-ynoic acid (Moya, 2a)
[0408] [.alpha.].sup.20.sub.D -19.0 (c 0.1, MeOH); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm: 2.47 (sextet, J=6.5 Hz, 1H), 2.19
(td, J=7.0, 2.5 Hz, 2H), 1.94 (br t, J=2.5 Hz, 1H), 1.73-1.67 (m,
1H), 1.54 (p, J=7.0, 2H), 1.49-1.41 (m, 3H), 1.19 (d, J=7.0 Hz,
3H); .sup.13C NMR (125 MHz, CDCl.sub.3) .delta. ppm: 183.1, 84.4,
68.5, 39.4, 33.0, 28.4, 26.4, 18.4, 17.0; HRMS (ESI) m/z calcd for
C.sub.9H.sub.15O.sub.2 [M+H].sup.+ 155.1067, found 155.1063.
(S)-2-methyloct-7-ynoic acid (2b)
[0409] (169.5 mg, 93%); [.alpha.].sup.20.sub.D 14.0 (c 0.1, MeOH);
.sup.1H (600 MHz, CDCl.sub.3) .delta. ppm: 2.50-2.45 (h, 1H), 2.20
(td, J=7.2, 3.0 Hz, 2H), 1.94 (t, J=3.0 Hz, 1H), 1.74-1.68 (m, 1H),
1.57-1.52 (m, 2H), 1.49-1.42 (m, 3H), 1.19 (d, J=7.0 Hz, 3H);
.sup.13C NMR (125 MHz, CDCl.sub.3) .delta. ppm: 182.5, 84.4, 68.5,
39.3, 33.1, 28.4, 26.4, 18.4, 17.0; HRMS (ESI) m/z calcd for
C.sub.9H.sub.1502 [M+H].sup.+ 155.1067, found 155.1064.
Preparation of Compounds 20a and 20b
##STR00162##
[0411] To the solution of N-Fmoc-L-Lys (Boc)-OH (5, 1.0 g, 2.13
mmol) and Meldrum's acid (6, 338.4 mg, 2.35 mmol) in anhydrous
CH.sub.2Cl.sub.2 (20 ml) at 0.degree. C. was added
4-dimethylaminopyridine (DMAP) (391 mg, 3.20 mmol) and EDCI-HCl
(530 mg, 2.77 mmol). After being stirred overnight at room
temperature, the reaction solution was diluted with EtOAc (60 ml),
washed by 5% citric acid (aq) (20 ml.times.4) and brine (20
ml.times.2), and the combined organic phase was then dried over
anhydrous MgSO.sub.4. The filtered EtOAc phase was refluxed under
heating for 30 min, then cooled down to room temperature and
evaporated to give crude intermediate 9a, which was used in the
next step without purification.
[0412] To a solution of the above crude 9a in the mixture of
diethyl ether (160 ml) and methanol (40 ml) was added
trimethylsilyldiazomethane (TMSCHN.sub.2) (4.4 ml, 8.79 mmol). The
resulting mixture was stirred overnight at room temperature, then
concentrated under reduced pressure. The residue was dissolved in
EtOAc (150 ml), washed with 5% citric acid (aq.) (30 ml.times.3),
saturate NaHCO.sub.3 (aq) (30 ml.times.2 and brine (30 ml), dried
over anhydrous MgSO.sub.4, and evaporated in vacuo. Product 20a was
obtained (558.0 mg, 50%) and 20b (18%) were obtained as white solid
after purification by silica gel column chromatography (eluted by
50-70% ethyl acetate in hexane).
(9H-fluoren-9-yl)methyl
(S)-2-(4-((tert-butoxycarbonyl)amino)butyl)-3-methoxy-5-oxo-2,5-dihydro-1-
H-pyrrole-1-carboxylate (20a)
[0413] [.alpha.].sup.20.sub.D 68.5 (c 0.1, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 7.77-7.74 (m, 4H), 7.40 (t, J=7.2 Hz,
2H), 7.33 (tt, J=7.1, 1.2 Hz, 2H), 5.09 (s, 1H), 4.58 (m, 2H), 4.47
(br s, 1H), 4.36 (dd, J=5.4, 3.0 Hz, 1H), 4.32 (t, J=6.6 Hz, 1H),
3.83 (s, 3H), 3.07-2.98 (m, 2H), 1.85-1.79 (m, 1H), 1.73-1.67 (m,
1H), 1.43 (s, 9H), 1.36-1.31 (m, 2H), 1.19-1.11 (m, 1H), 1.02-0.96
(m, 1H); .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm: 178.1,
168.9, 155.1, 151.1, 143.8, 143.7, 141.5, 141.4, 128.0, 127.9,
127.4, 125.4, 125.3, 120.1, 120.0, 94.5, 79.3, 68.1, 59.9, 58.8,
46.9, 40.5, 29.8, 28.7, 28.6, 19.6; HRMS (ESI) m/z calcd for
C.sub.29H.sub.35N.sub.2O.sub.6 [M+H].sup.+ 507.2490, found
507.2492.
[0414] Compound 20c,
##STR00163##
was prepared from 3',
##STR00164##
using the same procedure as used to prepare compounds 20a and
20b.
Preparation of Compound 3
##STR00165##
[0416] Piperidine (2.0 ml) was added to the solution of 4 (366.3
mg, 0.723 mmol) in MeCN (10 mL) at room temperature. After stirred
at the same temperature for 15 min, the reaction mixture was
concentrated and co-evaporated with toluene for 3 times. The
residue was purified by silica gel column chromatography (eluted by
EtOAc/hexane 1:1, then by 3-3.5% MeOH in CH.sub.2Cl.sub.2) to
afford 3 (180.0 mg, 88%) as white solid.
tert-butyl
(S)-(4-(3-methoxy-5-oxo-2,5-dihydro-1H-pyrrol-2-yl)butyl)carbam-
ate (3)
[0417] [.alpha.].sup.20.sub.D 10.0 (c 0.1, MeOH); .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. ppm: 6.30 (br s, 1H), 5.01 (s, 1H), 4.74
(t, J=6.0 Hz, 1H), 4.04 (dd, J=7.5, 4.0 Hz, 1H), 3.78 (s, 3H),
3.17-3.05 (m, 2H), 1.84-1.77 (m, 1H), 1.56-1.48 (m, 3H), 1.47-1.41
(m, 10H), 1.39-1.30 (m, 1H); .sup.13C NMR (125 MHz, CDCl.sub.3)
.delta. ppm: 178.5, 175.1, 156.2, 93.6, 78.9, 58.3, 57.5, 40.2,
31.4, 29.4, 28.5, 22.2; HRMS (ESI) m/z calcd for
C.sub.14H.sub.25N.sub.2O.sub.4 [M+H].sup.+ 285.1809, found
285.1808.
##STR00166##
were prepared using the same procedure used to prepare compound
3.
Preparation of Compounds 10a and 10b
##STR00167##
[0419] To the solution of 2a (29.1 mg, 0.1 mmol) in anhydrous
CH.sub.2Cl.sub.2 (3.0 ml) at 0.degree. C. was added
pentafluorophenol (40.1 mg, 0.22 mmol) in anhydrous
CH.sub.2Cl.sub.2 (0.5 ml), DMAP (2.4 mg, 0.02 mmol) and DCC (49.5
mg, 0.24 mmol). The resulting reaction mixture was stirred at room
temperature overnight and concentrated under reduced pressure. The
residue was stirred in cooled EtOAc (3.0 ml), and the suspending
solid was filtered off. The filtrate was evaporated in vacuo and
purified with silica gel column chromatography (eluted by 7% EtOAc
in hexane) to yield 10a (60.0 mg, 95%) as white solid. 10b was
synthesized from 2b following the same procedure.
Perfluorophenyl (R)-2-methyloct-7-ynoate (10a)
[0420] [.alpha.].sup.20.sub.D -27.5 (c 0.1, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 2.82 (h, J=6.6 Hz, 1H), 2.19 (td,
J=6.6, 3.0 Hz, 2H), 1.94 (br t, J=3.0 Hz, 1H), 1.87-1.81 (m, 1H),
1.65-1.51 (m, 5H), 1.34 (d, J=7.2 Hz, 3H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. ppm: 172.7, 142.1 (m), 140.5 (m), 138.8 (m),
137.2 (m), 125.4 (m), 84.2, 68.7, 39.4, 33.1, 28.3, 26.2, 18.4,
17.1; HRMS (ESI) m/z calcd for C.sub.15H.sub.14O.sub.2F.sub.5
[M+H].sup.+ 321.0908, found 321.0912.
Perfluorophenyl (S)-2-methyloct-7-ynoate (10b)
[0421] (84%); [.alpha.].sup.20.sub.D 14.0 (c 0.1, MeOH); .sup.1H
NMR (600 MHz, CDCl.sub.3) .delta. ppm: 2.82 (h, J=6.6 Hz, 1H), 2.23
(td, J=6.6, 2.4 Hz, 2H), 1.96 (t, J=2.4 Hz, 1H), 1.87-1.81 (m, 1H),
1.65-1.50 (m, 5H), 1.35 (d, J=7.2 Hz, 3H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. ppm: 172.7, 142.1 (m), 140.5 (m), 138.8 (m),
137.2 (m), 125.4 (m), 84.2, 68.7, 39.4, 33.1, 28.3, 26.2, 18.4,
17.1; HRMS (ESI) m/z calcd for C.sub.15H.sub.14O.sub.2F.sub.5
[M+H].sup.+ 321.0908, found 321.0915.
##STR00168##
were prepared using the same procedure used to prepare compounds
10a and 10b.
Preparation of Compounds 11a and 11b
##STR00169##
[0423] To a solution of 3 (16.9 mg, 0.06 mmol) in anhydrous THF
(1.0 ml) at -55.degree. C. was added n-BuLi (1.6 M in n-hexane)
(0.056 ml, 0.089 mmol), the solution was stirred the same
temperature for 30 min. Subsequently, the activated carboxylic acid
10a (28.5 mg, 0.089 mmol) in anhydrous THF (0.5 ml) was added
dropwise at -55.degree. C. The resulting reaction mixture was
stirred at the same temperature for 3 h and then room temperature
overnight. The reaction was quenched on the next day with saturate
NH.sub.4Cl (aq, 3.0 ml) and then extracted with EtOAc (5
ml.times.3). The combined organic phase was washed with saturate
NaHCO.sub.3 (aq, 5 ml.times.2) and brine (5 ml), dried over
anhydrous MgSO.sub.4, and purified by preparative TLC plate of
silica gel to yield corresponding product 11a (23.9 mg, 93%) as
white solid. 11b was obtained following the same procedure using
10b.
tert-butyl
(4-((S)-3-methoxy-1-((R)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-
-1H-pyrrol-2-yl)butyl)carbamate (11a)
[0424] [.alpha.].sup.20.sub.D 65.0 (c 0.1, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 5.04 (s, 1H), 4.65 (dd, J=5.4, 3.0
Hz, 1H), 4.51 (br s, 1H), 3.84 (s, 3H), 3.77 (sextet, J=6.6 Hz,
1H), 3.09-3.01 (m, 2H), 2.18 (td, J=6.6, 2.4 Hz, 2H), 2.08 (ddt,
J=13.8, 11.4, 5.4 Hz, 1H), 1.93 (t, J=3.0 Hz, 1H), 1.83 (dddd,
J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.71 (m, 1H), 1.57-1.49 (m,
2H), 1.48-1.39 (m, 14H), 1.21-1.14 (m, 1H), 1.12-1.06 (m, 4H);
.sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm: 179.1, 177.0,
170.0, 156.1, 94.3, 84.6, 79.3, 68.5, 59.2, 58.8, 40.5, 39.1, 33.7,
30.0, 28.8, 28.6, 28.5, 26.4, 20.0, 18.5, 18.3; HRMS (ESI) m/z
calcd for C.sub.23H.sub.37N.sub.2O.sub.5 [M+H].sup.+ 421.2697,
found 421.2692.
tert-butyl
(4-((S)-3-methoxy-1-((S)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-
-1H-pyrrol-2-yl)butyl)carbamate (11b)
[0425] (60%); [.alpha.].sup.20.sub.D 68.0 (c 0.1, MeOH); .sup.1H
NMR (600 MHz, CDCl.sub.3) .delta. ppm: 5.04 (s, 1H), 4.65 (dd,
J=5.4, 3.0 Hz, 1H), 4.51 (br s, 1H), 3.84 (s, 3H), 3.77 (sextet,
J=6.6 Hz, 1H), 3.07-3.04 (m, 2H), 2.17 (td, J=6.6, 2.4 Hz, 2H),
2.12 (ddt, J=13.8, 11.4, 5.4 Hz, 1H), 1.92 (t, J=3.0 Hz, 1H), 1.82
(dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.72 (m, 1H), 1.55-1.50
(m, 2H), 1.48-1.35 (m, 14H), 1.20-1.15 (m, 4H), 1.13-1.06 (m, 1H);
.sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm: 179.1, 177.0,
170.0, 156.0, 94.3, 84.7, 79.3, 68.3, 59.2, 58.8, 40.4, 39.3, 32.4,
29.9, 28.7, 28.7, 28.6, 26.6, 19.8, 18.5, 18.2; HRMS (ESI) m/z
calcd for C.sub.23H.sub.37N.sub.2O.sub.5 [M+H].sup.+ 421.2697,
found 421.2693.
##STR00170##
were prepared using the same procedure used to prepare compounds
11a and 11b.
Preparation of Compounds 11a-11d
##STR00171## ##STR00172## ##STR00173##
[0427] To the solution of compound 11a (2.1 mg, 0.005 mmol) in
CH.sub.2Cl.sub.2 (0.6 ml) was added TFA (0.3 ml) at 0.degree. C.
and the mixture was stirred the same temperature for 30 min. The
reaction was diluted with toluene (1 ml) and evaporated in vacuo (3
times) to produce crude 12a, which was used in next step without
purification. To the solution of crude 12a in anhydrous DMF (1.0
ml) was added 2a (0.8 mg, 0.005 mmol), EDCI.HCl (1.4 mg, 0.015
mmol), HOBt.H.sub.2O (1.2 mg, 0.008 mmol) and DIEA (3.0 .mu.L). The
reaction mixture was stirred at room temperature for 20 h and then
was evaporated in vacuo and purified by preparative TLC plate to
yield product 1a (1.8 mg, 79%) as white solid. 1b, 1c and 1d were
synthesized following the same procedure with corresponding
starting materials.
(R)--N-(4-((S)-3-methoxy-1-((R)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-1H--
pyrrol-2-yl)butyl)-2-methyloct-7-ynamide (Doscadenamide A, 1a)
[0428] [.alpha.].sup.20.sub.D 54.3 (c 0.07, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 5.47 (br t, J=6.0 Hz, 1H), 5.05 (s,
1H), 4.65 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.77 (sextet,
J=6.6 Hz, 1H), 3.27-3.21 (m, 1H), 3.17-3.12 (m, 1H), 2.20-2.17 (m,
4H), 2.14-2.11 (m, 1H), 2.07 (ddt, J=13.8, 11.4, 5.4 Hz, 1H), 1.94
(t, J=2.4 Hz, 1H), 1.93 (t, J=2.4 Hz, 1H), 1.85 (dddd, J=13.8,
11.4, 5.4, 3.0 Hz, 1H), 1.79-1.71 (m, 1H), 1.65-1.62 (m, 1H),
1.54-1.46 (m, 6H), 1.45-1.40 (m, 3H), 1.39-1.35 (m, 3H), 1.21-1.17
(m, 1H), 1.16-1.15 (m, 1H), 1.13-1.11 (m, 6H); .sup.13C NMR (150
MHz, CDCl.sub.3) .delta. ppm: 179.2, 177.0, 176.4, 170.0, 94.2,
84.7, 84.6, 68.5, 68.5, 59.2, 58.9, 41.8, 39.4, 39.1, 33.9, 33.7,
29.9, 29.6, 29.0, 28.5, 28.5, 26.7, 26.4, 20.4, 18.5, 18.4, 18.1,
16.3; HRMS (ESI) m/z calcd for C.sub.27H.sub.41N.sub.2O.sub.4
[M+H].sup.+ 457.3061, found 457.3058.
(S)--N-(4-((S)-3-methoxy-1-((R)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-1H--
pyrrol-2-yl)butyl)-2-methyloct-7-ynamide (1b)
[0429] (67%); [.alpha.].sup.20.sub.D 62.1 (c 0.07, MeOH); .sup.1H
NMR (600 MHz, CDCl.sub.3) .delta. ppm: 5.48 (br t, J=6.0 Hz, 1H),
5.05 (s, 1H), 4.65 (dd, J=6.0, 3.6 Hz, 1H), 3.85 (s, 3H), 3.77
(sextet, J=6.6 Hz, 1H), 3.26-3.20 (m, 1H), 3.19-3.13 (m, 1H),
2.20-2.16 (m, 4H), 2.15-2.11 (m, 1H), 2.07 (ddt, J=13.8, 11.4, 5.4
Hz, 1H), 1.94 (t, J=2.4 Hz, 1H), 1.93 (t, J=2.4 Hz, 1H), 1.85
(dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.72 (m, 1H), 1.65-1.63
(m, 1H), 1.54-1.47 (m, 6H), 1.45-1.41 (m, 3H), 1.39-1.35 (m, 3H),
1.22-1.18 (m, 1H), 1.18-1.14 (m, 1H), 1.12 (d, J=6.6 Hz, 3H), 1.12
(d, J=7.2 Hz, 3H); .sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm:
179.2, 177.0, 176.4, 170.0, 94.2, 84.7, 84.6, 68.5, 68.5, 59.2,
58.9, 41.7, 39.3, 39.1, 33.9, 33.7, 29.9, 29.6, 29.0, 28.5, 28.5,
26.8, 26.4, 20.4, 18.5, 18.4, 18.1, 16.3; HRMS (ESI) m/z calcd for
C.sub.27H.sub.41N.sub.2O.sub.4 [M+H].sup.+ 457.3061, found
457.3057.
(S)--N-(4-((S)-3-methoxy-1-((S)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-1H--
pyrrol-2-yl)butyl)-2-methyloct-7-ynamide (1c)
[0430] (82%); [.alpha.].sup.20.sub.D 95.0 (c 0.07, MeOH); .sup.1H
NMR (600 MHz, CDCl.sub.3) .delta. ppm: 5.45 (br t, J=5.4 Hz, 1H),
5.05 (s, 1H), 4.64 (dd, J=6.0, 3.0 Hz, 1H), 3.85 (s, 3H), 3.76
(sextet, J=7.2 Hz, 1H), 3.24-3.19 (m, 1H), 3.19-3.14 (m, 1H),
2.20-2.16 (m, 4H), 2.14-2.07 (m, 2H), 1.93 (t, J=3.0 Hz, 1H), 1.92
(t, J=3.0 Hz, 1H), 1.83 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H),
1.77-1.72 (m, 1H), 1.64-1.62 (m, 1H), 1.54-1.46 (m, 6H), 1.45-1.33
(m, 6H), 1.21-1.17 (m, 4H), 1.15-1.10 (m, 4H); .sup.13C NMR (150
MHz, CDCl.sub.3) .delta. ppm: 179.2, 177.0, 176.4, 170.0, 94.2,
84.7, 84.6, 68.5, 68.3, 59.2, 58.9, 41.7, 39.3, 39.2, 33.9, 32.5,
29.5, 28.9, 28.7, 28.5, 26.7, 26.6, 20.2, 18.5, 18.4, 18.1, 18.1;
HRMS (ESI) m/z calcd for C.sub.27H.sub.41N.sub.2O.sub.4 [M+H].sup.+
457.3061, found 457.3058.
(R)--N-(4-((S)-3-methoxy-1-((S)-2-methyloct-7-ynoyl)-5-oxo-2,5-dihydro-1H--
pyrrol-2-yl)butyl)-2-methyloct-7-ynamide (1d)
[0431] (62%); [.alpha.].sup.20.sub.D 60.0 (c 0.07, MeOH); .sup.1H
NMR (600 MHz, CDCl.sub.3) .delta. ppm: 5.44 (br t, J=6.0 Hz, 1H),
5.06 (s, 1H), 4.64 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.77
(sextet, J=6.6 Hz, 1H), 3.26-3.20 (m, 1H), 3.18-3.13 (m, 1H),
2.20-2.16 (m, 4H), 2.15-2.07 (m, 2H), 1.93 (t, J=3.0 Hz, 1H), 1.92
(t, J=3.0 Hz, 1H), 1.83 (dddd, J=13.8, 11.4, 6.0, 3.0 Hz, 1H),
1.77-1.72 (m, 1H), 1.64-1.62 (m, 1H), 1.54-1.46 (m, 6H), 1.44-1.34
(m, 6H), 1.21-1.16 (m, 4H), 1.16-1.10 (m, 4H); .sup.13C NMR (150
MHz, CDCl.sub.3) .delta. ppm: 179.2, 177.0, 176.4, 170.0, 94.3,
84.7, 84.6, 68.5, 68.3, 59.2, 58.9, 41.7, 39.3, 39.2, 33.9, 32.5,
29.9, 29.5, 29.0, 28.7, 28.5, 26.7, 26.6, 20.2, 18.5, 18.4, 18.1,
18.1; HRMS (ESI) m/z calcd for C.sub.27H.sub.41N.sub.2O.sub.4
[M+H].sup.+ 457.3061, found 457.3058.
[0432] The following compounds were prepared according to
aforementioned procedures:
##STR00174##
Isolation and Characterization of Doscadenamides B-I
[0433] Here we describe the identification of additional natural
doscadenamides (B-I) from the same cyanobacterium and the synthesis
of strategically diversified analogues to also probe the
structure-activity relationship (SAR) with respect to QS modulation
in several different gram-negative bacterial systems, and
interrogated the tentatively causative molecular interactions with
bacterial receptors. We then evaluated a subset of the focused
doscadenamide library in in cancer cells and successfully
established a synergistic activity with TRAIL in invasive triple
negative breast cancer cells, Careful chemical investigation of the
original extracts enabled to identification of several related
compounds that differed in the degree of unsaturation and
methylation pattern (FIG. 1). Rigorous 1D and 2D NMR analysis
coupled with HRMS and synthesis of selected family members
unambiguously established the structures of the additional
doscadenamides. Five pure structural analogs were isolated,
doscadenamides B-F (1b-1f, FIG. 1) as well as impure doscadenamides
G-J (1g-j, FIG. 1).
Doscadenamide B [1b; t.sub.R=10.1 min; 1.2 mg; white solid; HRESIMS
[M+H].sup.+ peak at m/z 455.2904, which suggested a molecular
formula C.sub.27H.sub.38N.sub.2O.sub.4. Doscadenamide C [(1c,
t.sub.R=13.7 min, 0.8 mg): white solid; HRESIMS [M+H].sup.+ peak at
m/z 459.3218, which suggested a molecular formula
C.sub.27H.sub.42N.sub.2O.sub.4. Doscadenamide D [(1d, t.sub.R=14.7
min, 0.8 mg): white solid; HRESIMS [M+H].sup.+ peak at m/z
459.3226, which suggested a molecular formula
C.sub.27H.sub.42N.sub.2O.sub.4. Doscadenamide E [(1e, t.sub.R=9.4
min, 0.1 mg): white solid; HRESIMS [M+H].sup.+ peak at m/z
445.3069, which suggested a molecular formula
C.sub.26H.sub.40N.sub.2O.sub.4. Doscadenamide F [(1f, t.sub.R=12.4
min, 0.8 mg): white solid; HRESIMS [M+H].sup.+ peak at m/z
461.3362, which suggested a molecular formula
C.sub.27H.sub.44N.sub.2O.sub.4. Doscadenamide G [(1f, t.sub.R=13.9
min, 0.3 mg): white solid; HRESIMS [M+H].sup.+ peak at m/z
445.3063, which suggested a molecular formula
C.sub.27H.sub.42N.sub.2O.sub.4. Doscadenamide I/J [(1i/1j,
t.sub.R=9.4 min, 0.5 mg, mixture), HRESIMS [M+H].sup.+ peak at m/z
442.2916, which suggested a molecular formula
C.sub.26H.sub.38N.sub.2O.sub.4.
[0434] We were not able to obtain pure doscadenamide G (1g, FIG.
1); therefore, based on our established synthetic route, we
synthesized the compound with the proposed structure (Scheme 1) and
compared its .sup.1H NMR spectrum with the isolated fraction,
confirming the dominant presence of doscadenamide H (1h). The
proposed doscadenamides H (1i) and J (1j) coeluted under various
HPLC conditions and were present in roughly equal abundance (1:1
mixture). To confirm our hypothesis, we synthesized the two
compounds, establishing the structures.
##STR00175##
Synthesis of a Strategic Focused Library to Probe the SAR
[0435] In addition to doscadenamides that are biosynthesized
through natural diversification, we aimed to prepare a
complementary compound set that possessed different degrees of
methylation and/or unsaturation in the carboxylic acid chains
(1j-1m), lacked one of the carboxylic acid chains (2a,b vs. 3a-3c),
exhibited different pyrrolinone configuration (1n) or transposed
pyrollinone functionalization (1o), as well as a cyclized version
(4a) (FIG. 1). We termed these new synthetic analogues
doscadenamides S4-S15 (FIG. 1). The diastereomers of 1a were
denominated as doscadenamides S1-S3 (FIG. 18).
[0436] To explore the contribution of each structural
characteristics in doscadenamide A (1a, FIG. 1) to the interaction
with its target and its biological activity, we synthesized most of
these structural analogs (doscadenamides F-J, 1f-1j) following
similar approach as previously described for 1a and exemplified for
the synthesis of doscadenamide H (1h, Scheme 1). Each target
compound can be obtained in three main steps: pyrrolinone ring
construction, carboxylic acid activation and amide/imide coupling.
For certain analogs (1f, 1k, 1l, 1m, 2b, 3b and 3c), we synthesized
additional carboxylic acids as specific building blocks to prepare
the desired products.
[0437] Our preliminary study indicates that the diastereomeric
doscadenamides S1-S3 behave similarly in modulating QS in
Pseudomonas aeruginosa. To perform the SAR study of doscadenamides
systematically, we selected several analogs to compose a focused
library for primary studies (FIG. 1, highlighted). To explore the
contribution of the unsaturated alkyne moiety to the bioactivity of
doscadenamide A (1a, FIG. 1), we included structural analogs with
two unsaturated alkene termini (1f, FIG. 1) and two saturated
carboxylic acid moieties (1k, FIG. 1). To further investigate the
significance of each side chain in doscadenamides, we included
structural analogs 2a and 3a-3c, possessing only one carboxylic
acid chain and with different degrees of unsaturation (3a-3c). To
confirm the importance of the linear structure of doscadenamide A
(1a, FIG. 1), the cyclic compound 4a was also included in the
library.
Total Synthesis of Doscadenamides
##STR00176##
[0439] Scheme 1 is a useful synthetic route for 1a. This synthetic
strategy was generalized to construct other analogs of (FIG. 1).
The reaction conditions were adjusted for the minority of
compounds. Various analogs of 5a as well as 6a (below) have been
synthesized using general methods, and the combination of building
blocks of 5a analogs with 6a analogs provided the series of target
molecules (1a-p, 2a, b, 3a-c, and 4a).
##STR00177##
##STR00178##
[0440] Similar to the synthesis of 5a and corresponding
intermediates, 5b, 5c and corresponding intermediates were
synthesized.
Synthesis of Compounds 10b
[0441] To the solution of N-Fmoc-D-Lys (Boc)-OH (1.0 g, 2.13 mmol)
and meldrum acid (338.4 mg, 2.35 mmol) in anhydrous
CH.sub.2Cl.sub.2 (20 ml) at 0.degree. C. was added
4-Dimethylaminopyridine (DMAP) (391 mg, 3.20 mmol) and EDCI-HCl
(530 mg, 2.77 mmol). After being stirred overnight at room
temperature, the reaction solution was diluted with EtOAc (60 ml),
washed by 5% citric acid (aq.) (20 ml.times.4) and brine (20
ml.times.2), dried over anhydrous MgSO.sub.4. The filtered EtOAc
phase was refluxed under heating for 30 min, then cooled down to
room temperature and evaporated to give crude intermediate 9b,
which was used in next step without purification.
[0442] To a solution of the above crude 9b in the mixture of
diethyl ether (160 ml) and methanol (40 ml) was added
trimethylsilyldiazomethane (TMSCHN.sub.2) (4.4 ml, 8.79 mmol). The
resulting mixture was stirred overnight at room temperature, then
concentrated under reduced pressure. The residue was dissolved in
EtOAc (150 ml), washed with 5% citric acid (aq.) (30 ml.times.3),
sat. NaHCO.sub.3 (30 ml.times.2 and brine (30 ml), dried over
anhydrous MgSO.sub.4, and evaporated in vacuo. Product 10b was
obtained after the crude was purified by chromatography column on
silica gel (eluted by 50-70% ethyl acetate in hexane).
Compound 10b (500.0 mg, 45.0% 2 Steps)
[0443] [.alpha.].sup.20.sub.D: -72.5 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 7.77-7.73 (m, 4H), 7.40 (t, J=7.2
Hz, 2H), 7.33 (t, J=7.2 Hz, 2H), 5.09 (s, 1H), 4.58 (m, 2H), 4.48
(br s, 1H), 4.36 (dd, J=5.4, 3.0 Hz, 1H), 4.33 (t, J=6.6 Hz, 1H),
3.83 (s, 3H), 3.08-2.98 (m, 2H), 1.85-1.79 (m, 1H), 1.71 (dddd,
J=14.4, 11.4, 4.8, 3.0 Hz, 1H), 1.44 (s, 9H), 1.38-1.31 (m, 2H),
1.19-1.11 (m, 1H), 1.03-0.96 (m, 1H) ppm. .sup.13C NMR (125 MHz,
CDCl.sub.3): .delta. 178.0, 168.8, 155.0, 151.1, 143.8, 143.7,
141.5, 141.4, 128.0, 127.9, 127.4, 125.4, 125.4, 120.1, 120.1,
94.5, 79.3, 68.1, 59.9, 58.8, 46.9, 40.5, 29.9, 28.7, 28.6, 19.6
ppm. HRMS (ESI) m/z calcd for C.sub.29H.sub.34N.sub.2O.sub.6
[M+H].sup.+ 507.2490, found 507.2492.
Compound 10c
[0444] In the methylation of pyrrolidine 9a previously.sup.2, both
4- and 2-carbonyls were methylated to give corresponding products
10a (50%) and 10c (18%), which were separable with chromatography
silica column. (18.2% from 9a). [.alpha.].sup.20.sub.D: -28.0 (c
0.1, MeOH). .sup.1H NMR (600 MHz, CDCl.sub.3): .delta. 7.78-7.77
(m, 2H), 7.58-7.55 (m, 2H), 7.43-7.40 (m, 2H), 7.35-7.31 (m, 2H),
4.82 (s, 1H), 4.73 (m, 2H), 4.45 (br s, 1H), 4.25 (t, J=4.8 Hz,
1H), 3.92 (s, 3H), 3.85-3.83 (m, 1H), 3.00-2.98 (m, 2H), 1.68-1.64
(m, 1H), 1.54-1.47 (m, 1H), 1.44 (s, 9H), 1.28-1.22 (m, 2H),
1.10-1.03 (m, 1H), 1.02-0.95 (m, 1H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 195.9, 176.6, 156.0, 149.5, 143.5, 143.4,
141.7, 141.6, 128.1, 128.1, 127.4, 127.4, 124.6, 124.5, 120.3,
120.3, 86.6, 79.2, 67.7, 66.1, 59.7, 47.0, 40.4, 29.9, 29.4, 28.6,
19.7 ppm. HRMS (ESI) m/z calcd for C.sub.29H.sub.34N.sub.2O.sub.6
[M+H].sup.+ 507.2490, found 507.2491.
Synthesis of Compounds 5b and 5c
[0445] Piperidine (2.0 ml) was added to the solution of 10b or 10c
(366.3 mg, 0.723 mmol) at room temperature. After stirred at the
same temperature for 15 min, the reaction solution was concentrated
and co-evaporated with toluene for 3 times. The residue was
purified by chromatography column on silica gel (eluted by
EtOAc/hexane 1:1, then by 3-3.5% MeOH in CH.sub.2Cl.sub.2) to
provide product 5b or 5c.
Compound 5b (93%)
[0446] [.alpha.].sup.20.sub.D: -21.5 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 6.23 (br s, 1H), 5.01 (s, 1H), 4.72
(t, J=6.0 Hz, 1H), 4.04 (dd, J=7.2, 3.6 Hz, 1H), 3.78 (s, 3H),
3.16-3.06 (m, 2H), 1.83-1.78 (m, 1H), 1.56-1.47 (m, 3H), 1.47-1.41
(m, 10H), 1.36-1.30 (m, 1H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 178.3, 174.5, 156.2, 93.7, 79.3, 58.5, 57.4,
40.3, 31.5, 29.9, 28.6, 22.1 ppm. HRMS (ESI) m/z calcd for
C.sub.14H.sub.24N.sub.2O.sub.4 [M+H].sup.+ 285.1809, found
285.1806.
Compound 5c (88%)
[0447] [.alpha.].sup.20.sub.D: -39.0 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.69 (br s, 1H), 4.66-4.64 (m, 2H),
3.89 (s, 3H), 3.75-3.73 (m, 1H), 3.13-3.07 (m, 2H), 1.91-1.85 (br
m, 1H), 1.66-1.60 (m, 1H), 1.53-1.47 (m, 2H), 1.44-1.37 (m, 11H)
ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 199.3, 181.6,
156.4, 80.4, 79.4, 63.5, 58.4, 40.1, 30.8, 30.0, 28.6, 22.2 ppm.
HRMS (ESI) m/z calcd for C.sub.14H.sub.24N.sub.2O.sub.4 [M+H].sup.+
285.1809, found 285.1807.
##STR00179##
Synthesis of 11a and 11b
[0448] To a solution of 7-Octenoic acid (Oea) (450 mg, 3.17 mmol)
and trimethylamine (Et.sub.3N) (0.56 mL, 4.13 mmol) in THF (18 mL)
at -20.degree. C. was added neat 2,2-trimethylacetyl chloride (0.43
mL, 3.49 mmol) dropwise over 20 min. The resulting mixture was
stirred at -20.degree. C. for 30 min and 0.degree. C. for another
30 min, then it was cooled to -78.degree. C. by dry ice-acetone. In
another reaction flask, n-butyllithium (n-BuLi) (1.6 M in n-hexane)
(2.0 ml, 3.17 mmol) was added dropwise to a solution of R- or
S-oxazolidione (841 mg, 4.75 mmol) in tetrahydrofuran at
-78.degree. C. The mixture was stirred at this temperature for 20
min and then transferred to the above solution of Oya in THF at
-78.degree. C. by cannula. The resulting mixture was stirred at
this temperature for 30 min, then it was allowed to warm to room
temperature and stirred for 1.5 h. The reaction was quenched with
saturate NH.sub.4Cl (aq) solution, extracted with EtOAc (45
mL.times.3), washed by 5% NaHCO.sub.3 solution, dried over
anhydrous MgSO.sub.4 and concentrated in vacuo. The residue was
purified by silica gel chromatography column (eluted by 15% ethyl
acetate in hexane) to give products 11a or 11b, corresponding to R-
or S-oxazolidione, respectively.
Compound 11a (451 mg, 78%)
[0449] [.alpha.].sup.20.sub.D: -97.0 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 7.35-7.32 (t m, 2H), 7.29-7.26 (t m,
1H), 7.22-7.20 (d m, 2H), 5.81 (ddt, J=17.5, 10.2, 6.6 Hz, 1H),
5.02-4.93 (m, 2H), 4.67 (ddt, J=10.8, 7.8, 3.0 Hz, 1H), 4.21-4.15
(m, 2H), 3.30 (dd, J=13.2, 3.0 Hz, 1H), 3.00-2.87 (m, 2H), 2.77
(dd, J=13.2, 9.6 Hz, 1H), 2.09-2.05 (m, 2H), 1.75-1.65 (m, 2H),
147-1.38 (m, 4H) ppm. .sup.13C NMR (125 MHz, CDCl.sub.3): .delta.
173.5, 153.6, 139.0, 135.5, 129.6, 129.1, 127.5, 114.6, 66.3, 55.3,
38.1, 35.6, 33.7, 28.8, 28.7, 24.2 ppm. HRMS (ESI) m/z calcd for
C.sub.18H.sub.23NO.sub.3 [M+H].sup.+ 302.1751, found 302.1748.
Compound 11b (907 mg, 95%)
[0450] [.alpha.].sup.20.sub.D: 85.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): .delta. 7.35-7.32 (t m, 2H), 7.29-7.26 (t m, 1H),
7.22-7.20 (d m, 2H), 5.81 (ddt, J=17.4, 10.2, 6.6 Hz, 1H),
5.02-4.93 (m, 2H), 4.67 (ddt, J=10.8, 7.8, 3.0 Hz, 1H), 4.21-4.15
(m, 2H), 3.30 (dd, J=13.2, 3.0 Hz, 1H), 3.00-2.87 (m, 2H), 2.77
(dd, J=13.2, 9.6 Hz, 1H), 2.09-2.05 (m, 2H), 1.76-1.65 (m, 2H),
147-1.38 (m, 4H) ppm. .sup.13C NMR (125 MHz, CDCl.sub.3): .delta.
173.5, 153.6, 139.0, 135.5, 129.6, 129.1, 127.5, 114.6, 66.3, 55.3,
38.1, 35.6, 33.7, 28.8, 28.7, 24.2 ppm. HRMS (ESI) m/z calcd for
C.sub.18H.sub.23NO.sub.3 [M+H].sup.+ 302.1751, found 302.1746.
Synthesis of Compounds 12a and 12b
[0451] To a solution of sodium bis(trimethylsilyl)amide (NaHMDS)
(2.0 M in THF) (1.11 ml, 2.21 mmol) in anhydrous THF (9 ml) at
-78.degree. C. was added compound 11a or 11b (609 mg, 2.02 mmol) in
THF (3.5 ml) under argon atmosphere. After the resulting reaction
solution was stirred at the same temperature for 30 min, neat MeI
(0.63 ml, 10.08 mmol) was added dropwise over 10 min. The reaction
mixture was quenched with saturate NH.sub.4Cl (aq.) (18 ml) after
it was stirred at -78.degree. C. for 20 h, then extracted with
ethyl acetate (25 ml.times.3), dried over anhydrous MgSO.sub.4, and
concentrated in vacuo. The residue was purified by chromatography
column of silica gel (eluted by 8% ethyl acetate in hexane) to give
product 12a or 12b, corresponding to 11a, 11b, respectively.
Compound 12a (566 mg, 89%)
[0452] [.alpha.].sup.20.sub.D: -70.0 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 7.34-7.31 (m, 2H), 7.29-7.26 (m,
1H), 7.22-7.21 (m, 2H), 5.79 (ddt, J=17.4, 10.2, 6.6 Hz, 1H),
5.01-4.92 (m, 2H), 4.68 (ddt, J=9.6, 7.8, 3.0 Hz, 1H), 4.21-4.16
(m, 2H), 3.70 (h, J=6.6 Hz, 1H), 3.27 (dd, J=13.2, 3.0 Hz, 1H),
2.77 (dd, J=13.2, 9.6 Hz, 1H), 2.05 (qt, J=7.0, 1.2 Hz, 2H),
1.78-1.72 (m, 1H), 1.46-1.36 (m, 3H), 1.36-1.29 (m, 2H), 1.22 (d,
J=6.6 Hz, 3H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta.
177.4, 153.2, 139.0, 135.5, 129.6, 129.1, 127.5, 114.6, 66.2, 55.5,
38.1, 37.8, 33.7, 33.4, 29.0, 26.8, 17.5 ppm. HRMS (ESI) m/z calcd
for C.sub.19H.sub.25NO.sub.3 [M+H].sup.+ 316.1907, found
316.1901.
Compound 12b (608 mg, 69%)
[0453] [.alpha.].sup.20.sub.D: 99.0 (c 0.1, MeOH). .sup.1H NMR (500
MHz, CDCl.sub.3): .delta. 7.33 (t, J=7.5 Hz, 2H), 7.27 (t, J=7.0
Hz, 1H), 7.21 (d, J=7.0 Hz, 2H), 5.79 (ddt, J=17.0, 10.0, 7.0 Hz,
1H), 5.00-4.92 (m, 2H), 4.70-4.65 (m, 1H), 4.21-4.15 (m, 2H), 3.70
(sextet, J=6.5 Hz, 1H), 3.26 (dd, J=13.5, 3.5 Hz, 1H), 2.77 (dd,
J=13.5, 9.5 Hz, 1H), 2.04 (q, J=7.0 Hz, 2H), 1.79-1.71 (m, 1H),
1.46-1.36 (m, 3H), 1.35-1.28 (m, 2H), 1.22 (d, J=7.0 Hz, 3H) ppm.
.sup.13C NMR (125 MHz, CDCl.sub.3): .delta. 177.4, 153.2, 139.0,
135.5, 129.6, 129.0, 127.5, 114.5, 66.1, 55.5, 38.0, 37.8, 33.7,
33.3, 29.0, 26.8, 17.5 ppm HRMS (ESI) m/z calcd for
C.sub.19H.sub.25NO.sub.3 (M+X).+-.[M+H].sup.+ 316.1907, found
316.1902.
Synthesis of Acid 6d, 6e
[0454] Hydrogen peroxide (30% in H.sub.2O) (0.7 ml, 6.12 mmol) was
added to the solution of 12a or 12b (480 mg, 1.52 mmol) in the
mixture of THF-H.sub.2O (10 ml-5 ml) at 0.degree. C. LiOH.H.sub.2O
(128.4 mg, 3.06 mmol) was added to the above reaction solution
after it was stirred at 0.degree. C. for additional 10 min. Then
the resulting reaction mixture was stirred at 0.degree. C. for 2 h
and additional 1 h at room temperature, then Na.sub.2SO.sub.3
(965.2 mg) was added. The quenched reaction was diluted with water
(25 ml) and EtOAc (25 ml). Ethyl acetate phase was separated and
the separated water phase was acidified with 1M aq. HCl (to pH 2)
and extracted EtOAc (25 ml.times.3). The combined EtOAc phase was
dried over anhydrous MgSO.sub.4, concentrated in vacuo and purified
by silica gel column chromatography (eluted by 17% ethyl acetate in
hexane). to give product 6d or 6e, corresponding to 12a, 12b,
respectively.
Acid 6d (246 mg, 92%)
[0455] [.alpha.].sup.20.sub.D: -23.0 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): 5.79 (ddt, J=16.8, 10.2, 6.6 Hz, 1H),
5.01-4.92 (m, 2H), .delta. 2.46 (sextet, J=7.2 Hz, 1H), 2.05 (qt,
J=8.4, 1.2 Hz, 2H), 1.72-1.67 (m, 1H), 1.47-1.32 (m, 5H), 1.18 (d,
J=6.6 Hz, 3H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta.
183.3, 138.9, 114.6, 39.5, 33.7, 33.5, 28.9, 26.7, 17.0 ppm. HRMS
(ESI) m/z calcd for C.sub.9H.sub.1602 [M+H].sup.+ 157.1223, found
157.1222.
Acid 6e (148.3, 62%)
[0456] [.alpha.].sup.20.sub.D: 13.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): 5.79 (ddt, J=16.8, 10.2, 6.6 Hz, 1H), 5.01-4.92
(m, 2H), .delta. 2.46 (sextet, J=7.2 Hz, xz1H), 2.05 (qt, J=8.4,
1.8 Hz, 2H), 1.72-1.66 (m, 1H), 1.47-1.32 (m, 5H), 1.18 (d, J=7.2
Hz, 3H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 183.4,
138.9, 114.6, 39.5, 33.7, 33.5, 28.9, 26.7, 17.0 ppm. HRMS (ESI)
m/z calcd for C.sub.9H.sub.1602 [M+H].sup.+ 157.1223, found
157.1219.
Synthesis of Acid 6g
[0457] Though acid 6g is a known and commercially available
compound, we prepared it conveniently from the available material
in lab. Acid 6d (33 mg, 0.211 mmol) was stirred under hydrogen
atmosphere in MeOH (2.0 mL) for 30 min, then the reaction mixture
was filtered and evaporated to provide acid 6g (30 mg, 90%).
Acid 6g (30 mg, 90%)
[0458] [.alpha.].sup.20.sub.D: -22.0 (c 0.15, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 2.45 (ddq, J=6.7 Hz, 1H), 1.71-1.65
(m, 1H), 1.45-1.40 (m, 1H), 1.34-1.27 (m, 8H), 1.17 (d, J=7.0 Hz,
3H), 0.88 (t, J=6.8 Hz, 3H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 182.9, 39.6, 33.8, 31.8, 29.3, 27.3, 22.8,
17.0, 14.2 ppm. HRMS (ESI) m/z calcd for C.sub.9H.sub.16O.sub.2
(M-H).sup.- 157.1229, found 157.1232.
##STR00180## ##STR00181## ##STR00182## ##STR00183##
##STR00184##
##STR00185## ##STR00186##
General Procedure for the Synthesis of PFP Ester 7 (Scheme 4)
[0459] To the solution of an acid 6 (0.1 mmol) in anhydrous
CH.sub.2Cl.sub.2 (3.0 mL) at 0.degree. C. was added
pentafluorophenol (40.1 mg, 0.22 mmol) in anhydrous
CH.sub.2Cl.sub.2 (0.5 mL), DMAP (2.4 mg, 0.02 mmol) and DCC (49.5
mg, 0.24 mmol). The resulting reaction mixture was stirred at room
temperature overnight and concentrated under reduced pressure. The
residue was stirred in cooled EtOAc (3.0 mL), and the suspending
solid was filtered off. The filtrate was evaporated in vacuo and
purified with silica gel column chromatography (eluted by 7% EtOAc
in hexane) to yield the corresponding PFP ester 7 as white
solid.
General Procedure for the Synthesis of 3a-e (Scheme 4)
[0460] To a solution of 5a (6.2 mg, 0.02 mmol) in anhydrous THF
(1.0 mL) at -55.degree. C. was added nBuLi (1.6 M in n-hexane)
(0.021 mL, 0.033 mmol), the solution was stirred the same
temperature for 30 min. Subsequently, the activated carboxylic
corresponding acid 7 (0.03 mmol) in anhydrous THF (0.5 mL) was
added dropwise at -55.degree. C. The resulting reaction mixture was
stirred at the same temperature for 3 h and then room temperature
overnight. The reaction was quenched on the next day with saturate
NH.sub.4Cl (aq, 3.0 mL) and then extracted with EtOAc (5
mL.times.3). The combined organic phase was washed with saturate
NaHCO.sub.3 (aq, 5 mL.times.2) and brine (5 mL), dried over
anhydrous MgSO.sub.4, and purified by preparative TLC plate of
silica gel to yield corresponding product 3 as white solid.
For the synthesis of 3f (Scheme 5a)), use 5b instead of 5a to
couple with PFP ester 7a following the above general procedure; For
the synthesis of 3g (Scheme 5b)), nBuLi was replaced by KHMDS (0.7
M in toluene) and use 5c instead of 5a to couple with PFP ester 7a
following the above general procedure.
General Procedure for the Synthesis of 1a-g (Schemes 4 and 5)
[0461] To the solution of corresponding 3 (0.010 mmol) in
CH.sub.2Cl.sub.2 (1.5 mL) was added TFA (0.75 mL) at 0.degree. C.
and the mixture was stirred the same temperature for 30 min. The
reaction was diluted with toluene (1 mL) and evaporated in vacuo (3
times) to produce an intermediate crude, which was used in next
step without purification. To the intermediate crude in anhydrous
DMF (1.0 mL) was added corresponding acid 6 (0.011 mmol), EDCI-HCl
(3.4 mg, 0.017 mmol), HOBt-H.sub.2O (2.8 mg, 0.018 mmol) and DIEA
(6.0 .mu.L). The reaction mixture was stirred at room temperature
for 20 h and then was evaporated in vacuo and purified by
preparative TLC plate to yield corresponding product doscadenamide
1 as white solid.
PFP Ester 7c (91.5%)
[0462] .sup.1H NMR (600 MHz, CDCl.sub.3): .delta. 2.68 (t, J=7.2
Hz, 2H), 2.23 (td, J=6.6, 2.4 Hz, 2H), 1.96 (t, J=3.0 Hz, 1H), 1.80
(p, J=7.8 Hz, 2H), 1.62-1.52 (m, 4H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 169.5, 142.1 (m), 140.5 (m), 138.8 (m), 137.2
(m), 125.3 (m), 84.2, 68.7, 33.3, 28.1, 28.0, 24.4, 18.4 ppm. HRMS
(ESI) m/z calcd for C.sub.14H.sub.11F.sub.5O.sub.2 [M+H].sup.+
307.0757, found 307.0760.
PFP Ester 7d (85%)
[0463] [.alpha.].sup.20.sub.D: -96.0 (c 0.03, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.81 (ddt, J=16.9, 10.2, 6.7 Hz,
1H), 5.03-4.94 (m, 2H), 2.81 (ddq, J=7.0 Hz, 1H), 2.10-2.06 (m,
2H), 1.86-1.80 (m, 1H), 1.63-1.56 (m, 1H) ppm, 1.48-1.40 (m, 4H),
1.33 (d, J=7.0 Hz). .sup.13C NMR (150 MHz, CDCl.sub.3): .delta.
172.8, 142.2 (m), 140.4 (m), 138.7 (m), 138.7, 137.2 (m), 125.4
(m), 114.7, 39.4, 33.7, 33.6, 28.8, 26.6, 17.1 ppm. HRMS (ESI) m/z
calcd for C.sub.15H.sub.15F.sub.5O.sub.2 (M+H).sup.+ 323.1070,
found 323.1068.
PFP Ester 7f (84%)
[0464] .sup.1H NMR (600 MHz, CDCl.sub.3): .delta. 5.81 (ddt,
J=17.4, 10.8, 6.6 Hz, 1H), 5.03-4.95 (m, 2H), 2.67 (t, J=7.2 Hz,
2H), 2.10-2.06 (m, 2H), 1.81-1.76 (m, 2H), 1.48-1.42 (m, 4H) ppm.
.sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 169.7, 142.1 (m), 140.5
(m), 138.9 (m), 138.7, 137.2 (m), 125.3 (m), 114.8, 33.6, 33.4,
28.5, 28.4, 24.8 ppm. HRMS (ESI) m/z calcd for
C.sub.14H.sub.13F.sub.5O.sub.2 [M+H].sup.+ 309.0914, found
309.0918.
PFP Ester 7g (90%)
[0465] [.alpha.].sup.20.sub.D: -25.0 (c 0.15, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 2.80 (ddq, J=7.0 Hz, 1H), 1.81 (dq,
J=13.6, 7.7, 1H), 1.62-1.57 (m, 1H), 1.42-1.36 (m, 2H), 1.36-1.28
(m, 9H), 0.89 (t, J=6.8 Hz, 3H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 172.9, 142.2 (m), 140.5 (m), 138.8 (m), 138.8,
137.2 (m), 125.5 (m), 114.7, 39.5, 33.7, 31.8, 29.2, 27.1, 22.7,
17.1, 14.2 ppm. HRMS (ESI) m/z calcd for C.sub.15H.sub.18F502
[M+H].sup.+ 325.1227, found 325.1222.
Doscadenamide S10 (3a)
[0466] [.alpha.].sup.20.sub.D 65.0 (c 0.1, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 5.04 (s, 1H), 4.65 (dd, J=5.4, 3.0
Hz, 1H), 4.51 (br s, 1H), 3.84 (s, 3H), 3.77 (sextet, J=6.6 Hz,
1H), 3.09-3.01 (m, 2H), 2.18 (td, J=6.6, 2.4 Hz, 2H), 2.08 (ddt,
J=13.8, 11.4, 5.4 Hz, 1H), 1.93 (t, J=3.0 Hz, 1H), 1.83 (dddd,
J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.71 (m, 1H), 1.57-1.49 (m,
2H), 1.48-1.39 (m, 14H), 1.21-1.14 (m, 1H), 1.12-1.06 (m, 4H);
.sup.13C NMR (150 MHz, CDCl.sub.3) .delta. ppm: 179.1, 177.0,
170.0, 156.1, 94.3, 84.6, 79.3, 68.5, 59.2, 58.8, 40.5, 39.1, 33.7,
30.0, 28.8, 28.6, 28.5, 26.4, 20.0, 18.5, 18.3; HRMS (ESI) m/z
calcd for C.sub.23H.sub.37N.sub.2O.sub.5 [M+H].sup.+ 421.2697,
found 421.2692.
Doscadenamide S11 (3b) (67%)
[0467] [.alpha.].sup.20.sub.D: 24.0 (c 0.04, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.78 (ddt, J=16.9, 10.2, 6.6 Hz,
1H), 5.04 (s, 1H), 5.00-4.90 (m, 2H), 4.65 (dd, J=5.7, 3.0 Hz, 1H),
4.48 (br s, 1H), 3.84 (s, 3H), 3.10-3.01 (m, 2H), 2.08 (ddt,
J=18.3, 9.9, 4.7 Hz, 1H), 2.03 (dt, J=7.1, 7.1 Hz, 2H), 1.83 (dddd,
J=14.2, 11.3, 5.4, 3.1 Hz, 1H), 1.74-1.72 (m, 1H), 1.67-1.63 (m,
1H), 1.45-1.37 (m, 14H), 1.36-1.29 (m, 2H), 1.20-1.14 (m, 1H),
1.13-1.08 (m, 4H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta.
179.1, 177.1, 170.0, 156.0, 139.0, 114.5, 94.3, 79.3, 59.2, 58.8,
40.5, 39.2, 34.1, 33.8, 29.9, 29.0, 28.8, 28.6, 26.7, 20.0, 16.3
ppm. HRMS (ESI) m/z calcd for C.sub.23H.sub.38N.sub.2O.sub.5
[M+Na].sup.+ 445.2678, found 471.2670.
Doscadenamide S12 (3c) (56%)
[0468] [.alpha.].sup.20.sub.D: 48.0 (c 0.033, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.04 (s, 1H), 5.00-4.90 (m, 2H),
4.66 (dd, J=5.6, 3.0 Hz, 1H), 4.48 (br s, 1H), 3.84 (s, 3H), 3.76
(ddq, J=6.7, 6.7, 6.7 Hz, 2H), 3.10-3.01 (m, 2H), 2.08 (ddt,
J=13.9, 10.9, 5.3 Hz, 1H), 1.83 (dddd, J=14.1, 11.3, 5.5, 3.1 Hz,
1H), 1.75-1.70 (m, 1H), 1.48-1.37 (m, 13H), 1.30-1.25 (m, 7H),
1.19-1.15 (m, 1H), 1.14-1.06 (m, 4H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 179.0, 177.2, 170.0, 156.0, 94.3, 79.3, 59.2,
58.8, 40.5, 39.2, 34.4, 32.0, 29.5, 28.8, 28.6, 27.2, 22.8, 20.0,
16.3, 14.2 ppm. HRMS (ESI) m/z calcd for
C.sub.23H.sub.40N.sub.2O.sub.5 [M+Na].sup.+ 447.2835, found
447.2825.
Compound 3d (71%)
[0469] [.alpha.].sup.20.sub.D: 60.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): .delta. 5.05 (s, 1H), 4.64 (dd, J=5.4, 3.0 Hz,
1H), 4.49 (br s, 1H), 3.85 (s, 3H), 3.08-3.03 (m, 2H), 2.99-2.88
(m, 2H), 2.20 (td, J=6.6, 2.4 Hz, 2H), 2.12 (ddt, J=13.8, 11.4, 5.4
Hz, 1.93 (t, J=3.0 Hz, 1H), 1.84 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz,
1H), 1.67 (p, J=7.2 Hz, 2H), 1.59-1.55 (m, 2H), 1.50-1.45 (m, 4H),
1.43 (s, 9H), 1.25-1.14 (m, 1H), 1.13-1.05 (m, 1H) ppm. .sup.13C
NMR (150 MHz, CDCl.sub.3): .delta. 179.1, 173.0, 170.3, 156.1,
94.2, 84.7, 79.3, 68.4, 59.2, 58.8, 40.5, 37.1, 30.0, 28.8, 28.6,
28.5, 28.4, 24.1, 20.0, 18.4 ppm. HRMS (ESI) m/z calcd for
C.sub.22H.sub.34N.sub.2O.sub.5 [M+H].sup.+ 407.2546, found
407.2549.
Compound 3e (62%)
[0470] [.alpha.].sup.20.sub.D: 65.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): .delta. 5.80 (ddt, J=17.4, 10.8, 6.6 Hz, 1H),
5.05 (s, 1H), 5.01-4.91 (m, 2H), 4.64 (dd, J=5.4, 3.0 Hz, 1H), 4.48
(br s, 1H), 3.85 (s, 3H), 3.09-3.02 (m, 2H), 2.98-2.87 (m, 2H),
2.12 (ddt, J=13.8, 11.4, 5.4 Hz, 1H), 2.08-2.03 (m, 2H), 1.84
(dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.66 (p, J=7.8 Hz, 2H),
1.46-1.36 (m, 15H), 1.20-1.14 (m, 1H), 1.12-1.05 (m, 1H) ppm.
.sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.1, 173.2, 170.3,
156.1, 130.1, 114.5, 94.2, 79.3, 59.2, 58.8, 37.2, 33.8, 29.9,
28.9, 28.9, 28.8, 28.6, 24.5, 20.0 ppm. HRMS (ESI) m/z calcd for
C.sub.22H.sub.36N.sub.2O.sub.5 [M+H].sup.+ 409.2697, found
409.2692.
Compound 3f (63%)
[0471] [.alpha.].sup.20.sub.D: -13.3 (c 0.06, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.06 (s, 1H), 4.65 (dd, J=5.4, 3.0
Hz, 1H), 4.48 (br s, 1H), 3.85 (s, 3H), 3.77 (sextet, J=6.6 Hz,
1H), 3.07-3.04 (m, 2H), 2.17 (td, J=7.2, 2.4 Hz, 2H), 2.12 (ddt,
J=13.8, 11.4, 5.4 Hz, 1H), 1.92 (t, J=3.0 Hz, 1H), 1.82 (dddd,
J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.73 (m, 1H), 1.56-1.49 (m,
2H), 1.48-1.36 (m, 14H), 1.19 (d, J=6.6 Hz, 3H), 1.17-1.14 (m, 1H),
1.12-1.07 (m, 1H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta.
179.1, 177.0, 170.0, 156.0, 94.3, 84.7, 79.3, 68.5, 68.3, 59.2,
58.8, 39.3, 32.4, 29.9, 28.7, 28.6, 26.6, 19.8, 18.5, 18.2 ppm.
HRMS (ESI) m/z calcd for C.sub.23H.sub.36N.sub.2O.sub.5 [M+H].sup.+
421.2697, found 421.2693.
Compound 3g (27%)
[0472] [.alpha.].sup.20.sub.D: -43.5 (c 0.1, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 4.90 (s, 1H), 4.52 (br s, 1H), 4.40
(dd, J=6.0, 3.0 Hz, 1H), 4.04 (s, 3H), 3.20 (h, J=6.6 Hz, 1H), 3.07
(br s, 2H), 2.21 (td, J=7.2, 2.4 Hz, 2H), 2.06-2.00 (m, 1H),
1.99-1.92 (m, 2H), 1.66-1.62 (m, 1H), 1.55-1.50 (m, 3H), 1.48-1.39
(m, 14H), 1.31-1.25 (m, 3H), 1.14 (d, J=7.2 Hz, 3H) ppm. .sup.13C
NMR (150 MHz, CDCl.sub.3): .delta. 179.1, 177.0, 170.0, 156.1,
94.3, 84.6, 79.3, 68.5, 59.2, 58.8, 40.5, 39.1, 33.7, 30.0, 28.8,
28.6, 28.5, 26.4, 20.0, 18.5, 18.3 ppm.
Doscadenamide F (10 (80%)
[0473] [.alpha.].sup.20.sub.D: 23.0 (c 0.21, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.78 (m, 2H), 5.43 (br t, J=5.8 Hz,
1H), 5.04 (s, 1H), 5.00-4.91 (m, 4H), 4.64 (dd, J=5.8, 3.0 Hz, 1H),
3.84 (s, 3H), 3.76 (ddq, J=6.8, 6.8, 6.8, 1H), 3.26-3.20 (m, 1H),
3.17-3.11 (m, 1H), 2.1 (ddq, J=7.2 Hz, 1H), 2.08-2.00 (m, 5H), 1.84
(dddd, J=14.2, 11.2, 5.5, 3.1 Hz, 1H), 1.77-1.71 (m, 1H), 1.64-1.59
(m, 1H), 1.51-1.43 (m, 2H), 1.43-1.30 (m, 8H), 1.29-1.24 (m, 2H),
1.23-1.13 (m, 2H), 1.11 (d, J=6.6 Hz, 3H), 1.11 (d, J=7.2 Hz, 3H)
ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 177.2,
176.6, 170.0, 139.0, 139.0, 114.5, 114.5, 94.2, 59.2, 58.8, 41.8,
39.3, 39.2, 34.3, 34.1, 33.8, 33.7, 29.6, 29.0, 29.0, 27.1, 26.8,
20.4, 18.1, 16.3 ppm. HRMS (ESI) m/z calcd for
C.sub.26H.sub.44N.sub.2O.sub.4 [M+H].sup.+ 461.33379, found
461.3369.
Doscadenamide H (1h) (69%)
[0474] [.alpha.].sup.20.sub.D: 54.3 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.80 (ddt, J=16.8, 10.2, 6.6 Hz,
1H), 5.44 (br t, J=6.0 Hz, 1H), 5.05 (s, 1H), 5.01-4.91 (m, 2H),
4.63 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.26-3.20 (m, 1H),
3.19-3.13 (m, 1H), 2.96-2.87 (m, 2H), 2.19-2.16 (m, 2H), 2.15-2.08
(m, 2H), 2.07-2.03 (m, 2H), 1.93 (t, J=2.4 Hz, 1H), 1.85 (dddd,
J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.68-1.63 (m, 3H), 1.54-1.45 (m,
4H), 1.44-1.34 (m, 7H), 1.23-1.17 (m, 1H), 1.15-1.09 (m, 4H) ppm.
.sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 176.4, 173.2,
170.3, 139.1, 114.5, 94.2, 84.6, 68.5, 59.1, 58.9, 41.8, 39.3,
37.2, 33.9, 33.8, 29.5, 28.9, 28.9, 28.8, 28.5, 26.7, 24.5, 20.3,
18.4, 18.1 ppm. HRMS (ESI) m/z calcd for
C.sub.26H.sub.40N.sub.2O.sub.4 [M+H].sup.+ 445.3061, found
445.3059.
Doscadenamide I (1i) (79%)
[0475] [.alpha.].sup.20.sub.D: 59.3 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.44 (br t, J=6.0 Hz, 1H), 5.05 (s,
1H), 4.63 (dd, J=6.0, 3.0 Hz, 1H), 3.85 (s, 3H), 3.26-3.20 (m, 1H),
3.19-3.13 (m, 1H), 2.99-2.89 (m, 2H), 2.20-2.16 (m, 4H), 2.15-2.08
(m, 2H), 1.93 (t, J=3.0 Hz, 2H), 1.85 (dddd, J=13.8, 11.4, 5.4, 3.0
Hz, 1H), 1.70-1.62 (m, 3H), 1.59-1.54 (m, 2H), 1.53-1.45 (m, 6H),
1.41-1.34 (m, 3H), 1.23-1.17 (m, 1H), 1.15-1.08 (m, 4H) ppm.
.sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 176.4, 173.1,
170.3, 94.2, 84.7, 84.6, 68.5, 68.4, 59.1, 58.9, 41.8, 39.3, 37.1,
33.9, 29.9, 29.6, 28.9, 28.5, 28.5, 28.4, 26.8, 26.7, 24.1, 20.3,
18.5, 18.4, 18.1 ppm. ppm. HRMS (ESI) m/z calcd for
C.sub.26H.sub.38N.sub.2O.sub.4 [M+H].sup.+ 443.2904, found
443.2900.
Doscadenamide J (1j) (100%)
[0476] [.alpha.].sup.20.sub.D: 52.9 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.48 (br t, J=6.0 Hz, 1H), 5.04 (s,
1H), 4.65 (dd, J=6.0, 3.0 Hz, 1H), 3.85 (s, 3H), 3.78 (h, J=6.6 Hz,
1H), 3.26-3.21 (m, 1H), 3.18-3.12 (m, 1H), 2.20-2.17 (m, 4H), 2.15
(t, J=7.2 Hz, 2H), 2.07 (ddt, J=13.8, 11.4, 5.4 Hz, 1H), 1.93 (t,
J=3.0 Hz, 1H), 1.84 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H),
1.78-1.73 (m, 1H), 1.65-1.62 (m, 1H), 1.56-1.51 (m, 4H), 1.50-1.47
(m, 2H), 1.46-1.40 (m, 6H), 1.23-1.14 (m, 2H), 1.12 (d, J=6.6 Hz,
3H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 177.0,
172.9, 170.0, 94.2, 84.7, 84.6, 68.5, 68.5, 59.2, 58.9, 39.5, 39.1,
36.8, 33.7, 29.5, 29.0, 28.5, 28.5, 28.3, 26.4, 25.4, 20.4, 18.5,
18.4, 16.3 ppm. HRMS (ESI) m/z calcd for
C.sub.26H.sub.38N.sub.2O.sub.4 [M+H].sup.+ 443.2904, found
443.2903.
Doscadenamide S4 (1k) (83%)
[0477] [.alpha.].sup.20.sub.D: 44.0 (c 0.12, MeOH); .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.78 (m, 2H), 5.43 (br t, J=5.8 Hz,
1H), 5.04 (s, 1H), 5.00-4.91 (m, 4H), 4.64 (dd, J=5.8, 3.0 Hz, 1H),
3.84 (s, 3H), 3.76 (ddq, J=6.8, 6.8, 6.8, 1H), 3.26-3.20 (m, 1H),
3.17-3.11 (m, 1H), 2.1 (ddq, J=7.2 Hz, 1H), 2.08-2.00 (m, 5H), 1.84
(dddd, J=14.2, 11.2, 5.5, 3.1 Hz, 1H), 1.77-1.71 (m, 1H), 1.64-1.59
(m, 1H), 1.51-1.43 (m, 2H), 1.43-1.30 (m, 8H), 1.29-1.24 (m, 2H),
1.23-1.13 (m, 2H), 1.11 (d, J=6.6 Hz, 3H), 1.11 (d, J=7.2 Hz, 3H)
ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 177.2,
176.6, 170.0, 139.0, 139.0, 114.5, 114.5, 94.2, 59.2, 58.8, 41.8,
39.3, 39.2, 34.3, 34.1, 33.8, 33.7, 29.6, 29.0, 29.0, 27.1, 26.8,
20.4, 18.1, 16.3 ppm. HRMS (ESI) m/z calcd for
C.sub.27H.sub.48N.sub.2O.sub.4 (M+H).sup.+ 465.3692, found
465.3685.
Doscadenamide S5 (1l) (77.5%)
[0478] [.alpha.].sup.20.sub.D: 102.1 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.80 (ddt, J=16.8, 10.2, 6.6 Hz,
1H), 5.45 (br t, J=5.4 Hz, 1H), 5.05 (s, 1H), 5.01-4.91 (m, 2H),
4.63 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.25-3.15 (m, 2H),
2.96-2.87 (m, 2H), 2.19-2.16 (m, 2H), 2.14-2.08 (m, 2H), 2.07-2.03
(m, 2H), 1.93 (t, J=2.4 Hz, 1H), 1.85 (dddd, J=13.8, 11.4, 5.4, 3.0
Hz, 1H), 1.68-1.63 (m, 3H), 1.54-1.45 (m, 4H), 1.44-1.34 (m, 7H),
1.23-1.16 (m, 1H), 1.16-1.08 (m, 4H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 179.1, 176.4, 173.2, 170.2, 139.1, 114.5,
94.2, 84.6, 68.5, 59.1, 58.9, 41.7, 39.2, 37.2, 33.9, 33.8, 29.5,
28.9, 28.9, 28.8, 28.5, 26.7, 24.5, 20.3, 18.4, 18.1 ppm. HRMS
(ESI) m/z calcd for C.sub.26H.sub.40N.sub.2O.sub.4 [M+H].sup.+
445.3061, found 445.3058.
Doscadenamide S6 (1m) (60%)
[0479] [.alpha.].sup.20.sub.D: 82.9 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.47 (br t, J=7.2 Hz, 1H), 5.05 (s,
1H), 4.63 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.25-3.15 (m, 2H),
2.99-2.89 (m, 2H), 2.21-2.16 (m, 4H), 2.14-2.08 (m, 2H), 1.94-1.93
(m, 2H), 1.85 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.69-1.63 (m,
3H), 1.59-1.56 (m, 2H), 1.56-1.42 (m, 6H), 1.41-1.35 (m, 3H),
1.22-1.14 (m, 1H), 1.13-1.09 (m, 4H) ppm. .sup.13C NMR (150 MHz,
CDCl.sub.3): .delta. 179.2, 176.4, 173.1, 170.3, 94.1, 84.7, 84.6,
68.5, 68.4, 59.2, 58.9, 41.8, 39.3, 37.1, 33.9, 29.5, 28.9, 28.5,
28.5, 28.5, 28.4, 26.7, 24.1, 20.2, 18.4, 18.1 ppm. HRMS (ESI) m/z
calcd for C.sub.26H.sub.38N.sub.2O.sub.4 [M+H].sup.+ 443.2904,
found 443.2903.
Doscadenamide S7 (1n) (58.1%)
[0480] [.alpha.].sup.20.sub.D: 42.9 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.79 (ddt, J=16.8, 10.2, 6.6 Hz,
1H), 5.46 (br t, J=5.4 Hz, 1H), 5.05 (s, 1H), 5.01-4.92 (m, 2H),
4.65 (dd, J=6.6, 3.0 Hz, 1H), 3.85 (s, 3H), 3.80-3.75 (m, 1H),
3.26-3.20 (m, 1H), 3.18-3.12 (m, 1H), 2.18 (td, J=7.2, 2.4 Hz, 2H),
2.14 (t, J=7.2 Hz, 2H), 2.10-2.02 (m, 3H), 1.93 (t, J=2.4 Hz, 1H),
1.85 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.78-1.73 (m, 1H),
1.65-1.60 (m, 5H), 1.57-1.49 (m, 3H), 1.49-1.37 (m, 7H), 1.35-1.29
(m, 2H), 1.23-1.14 (m, 2H), 1.12 (d, J=6.6 Hz, 3H) ppm. .sup.13C
NMR (150 MHz, CDCl.sub.3): .delta. 179.2, 177.0, 173.1, 170.0,
138.9, 114.6, 94.2, 84.7, 68.5, 59.2, 58.9, 39.5, 39.1, 36.9, 33.7,
29.5, 29.0, 28.9, 28.7, 28.5, 26.4, 25.8, 20.4, 18.5, 16.3 ppm.
HRMS (ESI) m/z calcd for C.sub.26H.sub.40N.sub.2O.sub.4 [M+H].sup.+
445.3061, found 445.3059.
Doscadenamide S13 (1o) (66%)
[0481] [.alpha.].sup.20.sub.D: -40.0 (c 0.09, MeOH).). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.42 (br t, J=6.0 Hz, 1H), 5.06 (s,
1H), 4.64 (dd, J=5.4, 3.0 Hz, 1H), 3.85 (s, 3H), 3.77 (sextet,
J=6.6 Hz, 1H), 3.25-3.20 (m, 1H), 3.19-3.14 (m, 1H), 2.20-2.16 (m,
4H), 2.14-2.08 (m, 2H), 1.93 (t, J=2.4 Hz, 1H), 1.92 (t, J=2.4 Hz,
1H), 1.83 (dddd, J=13.8, 11.4, 5.4, 3.0 Hz, 1H), 1.77-1.72 (m, 1H),
1.65-1.61 (m, 1H), 1.53-1.46 (m, 6H), 1.45-1.34 (m, 3H), 1.32-1.28
(m, 3H), 1.18 (d, J=7.2 Hz, 3H), 1.16-1.13 (m, 2H), 1.12 (d, J=6.6
Hz, 3H) ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 179.2,
177.0, 176.4, 170.0, 94.2, 84.7, 84.6, 68.5, 68.3, 59.2, 58.9,
41.7, 39.3, 39.3, 33.9, 32.5, 32.1, 29.5, 29.2, 29.0, 28.7, 28.5,
26.8, 26.6, 20.2, 18.5, 18.4, 18.1, 18.1 ppm. HRMS (ESI) m/z calcd
for C.sub.27H.sub.40N.sub.2O.sub.4 [M+H].sup.+ 457.3061, found
457.3056.
Doscadenamide S14 (1p) (38%)
[0482] [.alpha.].sup.20.sub.D: -41.4 (c 0.07, MeOH). .sup.1H NMR
(600 MHz, CDCl.sub.3): .delta. 5.59 (br t, 1H), 4.90 (s, 1H), 4.40
(dd, J=6.0, 3.6 Hz, 1H), 4.04 (s, 3H), 3.27-3.15 (m, 3H), 2.22-2.13
(m, 5H), 2.04-1.93 (m, 4H), 1.79-1.72 (m, 1H), 1.67-1.60 (m, 1H),
1.56-1.47 (m, 6H), 1.46-1.40 (m, 3H), 1.39-1.35 (m, 3H), 1.33-1.28
(m, 2H), 1.14 (d, J=6.6 Hz, 3H), 1.11 (d, J=6.6 Hz, 3H) ppm.
.sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 196.9, 176.5, 175.2,
174.0, 86.6, 84.7, 84.4, 68.7, 68.5, 65.7, 59.9, 41.7, 40.3, 39.1,
33.9, 29.4, 29.2, 28.6, 28.4, 26.8, 26.5, 20.5, 19.3, 18.5, 18.4,
18.1, 16.9 ppm. HRMS (ESI) m/z calcd for
C.sub.27H.sub.40N.sub.2O.sub.4 [M+H].sup.+ 457.3061, found
457.3055.
##STR00187##
Synthesis 2a or 2b (Scheme 6)
[0483] To the solution of compound 5a (4.5 mg, 0.016 mmol) in
CH.sub.2Cl.sub.2 (0.8 mL) was added TFA (0.4 mL) at 0.degree. C.
and the mixture was stirred the same temperature for 30 min. The
reaction residue was diluted with toluene (1 mL) and evaporated in
vacuo (3 times) to produce an intermediate crude that was used in
next step without purification. To the solution of this
intermediate crude in anhydrous DMF (1.0 mL) was added
corresponding acid 6 (2.5 mg, 0.016 mmol), EDCI-HCl (4.6 mg, 0.024
mmol), HOBt-1420 (3.9 mg, 0.025 mmol) and DIEA (8.4 .mu.L). The
reaction mixture was stirred at room temperature for 20 h and then
was evaporated in vacuo and purified by preparative TLC plate to
yield corresponding product 2 as white solid.
Doscadenamide S8 (2a) (33% 2 Steps)
[0484] [.alpha.].sup.20.sub.D: -5.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): .delta. 5.93 (br s, 1H), 5.47 (br t, J=5.4 Hz,
1H), 5.00 (s, 1H), 4.05 (dd, J=7.2, 3.6 Hz, 1H), 3.79 (s, 3H),
3.32-3.27 (m, 1H), 3.24-3.18 (m, 1H), 2.20-2.13 (m, 3H), 1.93 (t,
J=2.4 Hz, 1H), 1.86-1.80 (m, 1H), 1.67-1.64 (m, 1H), 1.58-1.49 (m,
5H), 1.44-1.36 (m, 4H), 1.36-1.28 (m, 1H), 1.14 (d, J=7.2 Hz, 3H)
ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 178.3, 176.7,
174.5, 93.7, 84.6, 68.5, 58.5, 57.4, 41.7, 39.0, 31.3, 29.6, 28.5,
26.8, 22.0, 18.5, 18.2 ppm. HRMS (ESI) m/z calcd for
C.sub.18H.sub.28N.sub.2O.sub.3 [M+H].sup.+ 321.2173, found
321.2169.
Doscadenamide S9 (2b) (43% 2 Steps)
[0485] [.alpha.].sup.20.sub.D: 17.0 (c 0.1, MeOH). .sup.1H NMR (600
MHz, CDCl.sub.3): .delta. 6.19 (br s, 1H), 5.68 (br t, J=5.4 Hz,
1H), 4.99 (s, 1H), 4.05 (dd, J=7.8, 4.2 Hz, 1H), 3.79 (s, 3H),
3.32-3.27 (m, 1H), 3.23-3.18 (m, 1H), 2.20-2.13 (m, 3H), 1.93 (t,
J=3.0 Hz, 1H), 1.85-1.80 (m, 1H), 1.67-1.64 (m, 1H), 1.57-1.48 (m,
5H), 1.44-1.36 (m, 4H), 1.36-1.29 (m, 1H), 1.13 (d, J=7.2 Hz, 3H)
ppm. .sup.13C NMR (150 MHz, CDCl.sub.3): .delta. 178.4, 176.7,
174.6, 93.7, 84.6, 68.5, 58.5, 57.4, 41.7, 39.0, 33.9, 31.3, 29.5,
28.5, 26.8, 22.1, 18.5, 18.1 ppm. HRMS (ESI) m/z calcd for
C.sub.18H.sub.28N.sub.2O.sub.3 [M+H].sup.+ 321.2173, found
321.2169.
##STR00188##
[0486] To a solution of 1a (1.0 mg, 0.0022 mmol) in anhydrous THF
(0.5 mL) was added pyridine (0.25 mL), Cu(OAc).sub.2 (23.8 mg,
0.131 mmol) and CuI (6.3 mg, 0.033 mmol) sequentially. The reaction
was stirred at room temperature overnight and the purification with
preparative TLC to provide product 4a (0.8 mg, 80%).
Doscadenamide S15 (4a)
[0487] [.alpha.].sup.20.sub.D 14.0 (c 0.07, MeOH); .sup.1H NMR (600
MHz, CDCl.sub.3) .delta. ppm: 5.61 (br t, J=5.90 Hz, 1H), 5.06 (s,
1H), 4.69 (dd, J=5.8, 3.0 Hz, 1H), 3.90 (ddt, J=6.8, 5.0, 3.3 Hz,
1H), 3.86 (s, 3H), 3.28-3.19 (m, 2H), 2.31-2.24 (m, 4H), 2.19 (dqd,
J=10.3, 6.7, 3.9, 1H), 2.10 (ddt, J=14.1, 10.9, 5.2, 1H), 1.90
(dddd, J=14.3, 11.5, 5.6, 3.2 Hz, 1H), 1.80 (dtd, J=13.5, 9.3, 5.2
Hz, 1H), 1.67-1.65 (m, 1H), 1.52-1.42 (m, 9H), 1.32-1.28 (m, 3H),
1.19-1.15 (m, 2H), 1.12 (d, J=6.8, 6H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. ppm: 179.2, 176.9, 176.2, 170.1, 94.2, 77.7,
77.7, 66.1, 65.4, 59.1, 58.9, 41.7, 41.2, 39.6, 38.8, 34.0, 29.7,
29.5, 28.2, 27.1, 26.3, 26.1, 20.6, 19.4, 19.2, 18.6, 17.2; HRMS
(ESI) m/z calcd for C.sub.27H.sub.38N.sub.2O.sub.4(M+H).sup.+
455.2910, found 455.2899.
Quorum Sensing Signaling
[0488] Preliminary screening results indicated that 1a can activate
the 3-oxo-C12-HSL-responsive reporter plasmid pSB1075, a plasmid
encoding LasR and containing a light-producing luxCDABE cassette
expressed in E. coli (FIG. 15) [Winson, M. K.; Swift, S.; Fish, L.;
Throup, J. P.; Jorgensen, F.; Chhabra, S. R.; Bycroft, B. W.;
Williams, P.; Stewart, G. S., FEMS Microbiol. Lett. 1998, 163 (2),
185-192]. However, 1a was not able to activate the related reporter
pTIM5319, which is identical to pSB1075 but lacks the AHL-binding
site LasR (FIG. 16), which suggests that 1a exerts the QS
activating activity via the AHL-binding site [Kwan, J. C.; Meickle,
T.; Ladwa, D.; Teplitski, M.; Paul, V.; Luesch, H., Mol. Biosyst.
2011, 7 (4), 1205-1216]. To validate the activating activity of 1a,
its effect on wild-type P. aeruginosa was tested. As shown in FIG.
17, after treatment with 1a, the production and expression of the
secreted QS pigment pyocyanin was elevated. For comparison, the
other three diastereomers 1b, 1c and 1d were also included in the
test. Such superagonists are expected to artificially regulate
virulence factor production such as pyocyanin and activate QS at
lower bacterial cell populations, thus to stimulate the host immune
system to clear the infection when fewer bacterial cells are
present [Galloway, W.; Hodgkinson, J.; Bovvden, S.; Welch, M.;
Spring, D., Trends Microbiol. 2012, 20 (9), 449-458].
QS Modulation by Doscadenamides in Pseudomonas aeruginosa and LasR
Molecular Interaction Studies
[0489] Doscadenamide A (1a) activates QS, which was established
using the 3-oxo-C12-HSL-responsive reporter plasmid pSB1075, a
plasmid encoding LasR (the C12 HSL receptor in Pseudomonas
aeruginosa) and containing a light-producing luxCDABE cassette
expressed in Escherichia coli. The activity was abolished in a
strain (pTIM5319) that lacked the AHL domain but was otherwise
identical.
[0490] To follow up our previous study regarding the QS modulatory
activity of doscadenamide A (1a) on wild-type P. aeruginosa and
extend reporter gene assay studies, we examined the QS activation
of different doscadenamides using a QS system deficient strain,
PAO-JP1. PAO-JP1 is a P. aeruginosa mutant with a lasl deletion
that cannot produce C12. To investigate the activating effect of
doscadenamide A (1a) on P. aeruginosa without interference from the
C12 produced by P. aeruginosa itself, we examined the pyocyanin
production of P. aeruginosa mutant PAO-JP1 after treatment with
doscadenamide A (1a) and selected structural analogues for 6 h at
100 .mu.M.
[0491] Compounds selected for this bioassay possessed a different
number of carboxylic acid chains (1a vs 2a vs 3a), different
degrees of unsaturation (1a/f/k vs 3a-c). As shown in FIG. 3B, when
normalized for cell number, these doscadenamides (FIG. 1) can all
activate the pyocyanin production of PAO-JP1 but to a different
extent at the concentration of 100 .mu.M. Doscadenamide A (1a)
induced 3-fold QS activation, similarly to the positive control
C12. Compound 2a, which possesses only one chain (Moya1), exhibits
minimal QS-activating effect, while 3a-c with acylation of the
amide core showed the most potent activation, 3-fold, 4-fold and
5-fold, indicating that this chain is predominantly responsible for
the QS agonist activity in P. aeruginosa. The degree of
unsaturation appears to play a minor role in chain-dependent
fashion as the trends for 1a/f/k vs 3a-c were opposite, possibly
indicating the dependence of both chains to the overall net effect.
The overall trend was the same even without normalization for
bacterial cell count, except that 2a did not show any level of
activation, further underscoring the usefulness of Moya2
(regardless of degree of saturation) for QS activation.
QS Modulation by Doscadenamides in Other Gram-Negative Bacteria:
Vibrio harveyi
[0492] We then aimed to determine if doscadenamides modulate QS in
other Gram-negative bacteria as well. To extend our investigation
of doscadenamides on QS modulation, we adopted a bioluminescent
marine bacterium, Vibrio harveyi, as our model system to examine
the effect of doscadenamides on modulating the bioluminescence
production by V. harveyi. V. harveyi has been reported to be
responsive to bacterial QS activator
N-(3-oxododecanoyl-L-homoserine lactone (C12) and thus a robust
model for quorum sensing related research.
[0493] Most Gram-negative bacteria possess QS circuits similar to
the marine symbiotic bioluminescent bacterium Vibrio fischeri, the
QS system of which has been studied the most. As the V. fischeri
population grows, it produces and releases autoinducer molecules to
accumulate and thus eliciting bioluminescence. There are two
proteins, LuxI and LuxR, in V. fischeri to regulate QS signaling
pathway. LuxI regulates the production of the autoinducer,
N-(3-oxohexanoyl)-homoserine lactone (C6). LuxR binds to the
autoinducer and activates the transcription of luciferase enzymes
for bioluminescence production. In P. aeruginosa, two pairs of
LuxI/LuxR homologues, namely LasI/LasR and Rh1I/Rh1R, exist to
regulate the QS signaling pathway. For these two signaling systems,
the Rh1I/Rh1R system is subordinate to the LasI/LasR system. Two
acylhomoserine lactones (AHLs), C12 and N-butyryl-homoserine
lactone (C4), are required to trigger the expression of Rh1R and
its downstream target genes including virulence factor production;
while the expression of LasR is independent of the AHL-mediated QS
signaling pathway, thus making LasR a representative target for QS
inhibition. Compared to V. fischeri and P. aeruginosa, V. harveyi
contains a more complex QS cascade involving three parallel
regulating QS-signaling pathways and responds to three different
autoinducers. Even though the homologous LuxI/LuxR system of V.
fischeri has never been identified in V. harveyi, V. harveyi can
produce and detect autoinducer 1 (AI-1), N-(3-hydroxybutanoyl)
homoserine lactone, which shares significant similarity to C12 and
C4. Tyr 56 and Asp73 are conserved in both LasR and LuxR systems.
Therefore, we adopted V. harveyi as a practical model to understand
and validate the QS modulating activity of doscadenamide A (1a) and
its analogs (FIG. 1).
[0494] Almost all compounds activated QS in this biological context
to a certain extent (FIG. 19C), with 1a and 3a activating across
the concentration range tested (10-200 .mu.M). However, 3c only had
a minor activating trend and for 1f and 1j the activities at
.ltoreq.100 .mu.M were also reduced, suggesting that increased
saturation of the terminal units decreases activity for both
chains. Notably, 2a was not able to activate the bioluminescent
response in V. harveyi but, instead, consistently inhibited QS
signaling in V. harveyi, whereas the same compound was essentially
inactive in P. aeruginosa. In both cases, 2a was not able to
activate QS, providing common ground in the two Gram-negative
bacteria. The same trend in V. harveyi was observed without
normalization for cell number, indicating that the differential
effect is not an artifact of normalization.
[0495] These results indicate that both acylations in doscadenamide
A (1a) are involved in the QS modulating activity, with partially
overlapping but also bacteria context-specific trends. Both chains
contribute differently to the final biological activity output,
even in opposing or interacting fashion, suggesting a more complex
interaction between doscadenamide A (1a) and its molecular targets
in various bacterial systems, potentially regulating their
respective activity.
Synergy with TRAIL in MDA-MB-231 Breast Cancer Cells
[0496] TRAIL has been regarded as an attractive therapeutic
anticancer agent; however, many cancer cells have developed
resistance to TRAIL and impacted the treatment efficacy. Given that
C12 was reported to exhibit synergistic effect on TRAIL-induced
apoptosis in cancer cells, we examined the potential synergistic
effect of doscadenamide A (1a) and its analogs on sensitizing
cancer cells. First, we generated the dose response of TRAIL on
MDA-MB-231 breast cancer cells, which were partially responsive to
TRAIL, 50% cell viability remaining at 500 ng/mL (FIG. 20A), and
selected 20 ng/mL as our test concentration, under which TRAIL
would only induce a small fraction (approximately 20%) of cells to
undergo apoptotic death. MDA-MB-231 cells were pretreated with C12,
doscadenamides or solvent control for 3 h and then stimulated with
TRAIL for 24 h (FIGS. 20B and 20C). We measured cell viability and
calculated the A Bliss independence of each compound with TRAIL.
Most analogs exhibited synergistic effect with TRAIL, except 2a
(FIG. 1), and among all the analogs, the "saturated" analogs 1k and
3c (FIG. 1) behaved as the best agents to sensitize MDA-MB-231
cells (FIG. 20D). To further investigate the pro-apoptotic effect
of doscadenamide A (1a), the PARP cleavage was also analyzed using
Western blot (FIG. 20E) to indicate programmed cell death. There
was a clear decrease in cell viability and an evident presence of
cleaved PARP in cell extracts after co-treatment; therefore, the
doscadenamide A (1a) and compounds of the disclosure can
effectively improve TRAIL-induced apoptosis in MDA-MB-231 cells.
Furthermore, our SAR study reveals that the less the unsaturation
degree the compound possesses, the more effectively it acts on
ameliorating TRAIL resistance and the one side chain Moya2 plays a
useful role in exhibiting bioactivities, which generally correlated
with our QS results, in that the side chain Moya2 was useful for
activity since 2a showed no effect on cell viability (FIGS. 20B and
20C) or effect on PARP cleavage. Compound 3c showed even enhanced
activity compared with the doubly acylated versions, including
parent compound, doscadenamide A (1a). As expected, the cyclized
version 4a was completely inactive in these assays. Interestingly,
compared to doscadenamide A (1a), the analogs with less
unsaturation (1f, 1k, 3b and 3c) exhibit improved cytoxicity.
Biological Experimental Procedures
Quorum Sensing Reporter Assays with Doscadenamide a (1a)
[0497] Doscadenamide A (1a) in a dose-response manner and EtOH
control were added to the corresponding wells in a 96-well plate
and the solvent was allowed to evaporate. Then 100 .mu.L of an
overnight culture of E. coli expressing plasmid wild-type pSB1075
(a luxCDABE reporter construct encoding LasR) or an overnight E.
coli culture expressing reporter pSB1075 mutant (Y56F, W60F or
D73N), was added to each well. The plate was incubated at
37.degree. C. for 6 h before the measurement of luminescence.
Pyocyanin Quantification in P. aeruginosa Strain PAO-JP1
[0498] An overnight culture of P. aeruginosa strain PAO-JP1 was
diluted 10-fold before transferred to another culture tube
containing 890 .mu.L LB broth and 10 .mu.L testing compound (10
.mu.M final concentration) or EtOH control and incubated at
37.degree. C. with shaking for 6 h. The culture was then spun down
for 10 min at the maximum speed, and the supernatant was collected
and filtered using 0.2 .mu.M Eppendorf filters. 500 .mu.L of the
sterile supernatant were added to 500 .mu.L CHCl.sub.3 in an
Eppendorf tube. Tube shaking allowed for the extraction of
pyocyanin in the CHCl.sub.3 layer. This layer was then added to 150
.mu.L of 0.2 N HCl in another Eppendorf tube. After shaking, the
aqueous layer and the organic layer were separated. 100 .mu.L of
the aqueous layer were transferred to a clear bottomed 384-well
plate and the absorbance was measured at 385 nm to quantify the
pyocyanin production. Data was analyzed using GraphPad Prism 5
software.
Bioluminescence Modulation in V. harveyi
[0499] Bioluminescent V. harveyi BB120 strain (wild-type) was
cultivated in AB media overnight (15-16 h) at 30.degree. C. (0D600
is about 2.0-2.5). The overnight cultivated strain 2 .mu.L was
diluted to 10 mL by AB media (0D600<0.1). 100 .mu.L the diluted
BB120 strain was distributed to each well of 96-well white plate.
The solution of each tested compound in DMSO (0.5 .mu.L) at
different concentration was added to BB120 cultures on plate.
Triplicate was conducted for each compound and each concentration.
After addition, bacterial cultures were incubated at 30.degree. C.
for 7 hours. Then the emitted luminescence and OD600 was measured
by BioTek Synergy H.sub.1 plate reader. AI-1 and DMSO were used as
positive control and negative control, respectively.
Synergistic Effect Evaluation of Doscadenamide Analogs with
TRAIL
[0500] MDA-MB-231 cells were seeded in 12-well plates at a density
of 100,000 cells per well. The next day, cells were pretreated with
doscadenamide analogs (50 .mu.M and 25 .mu.M), positive controls
(C12) or solvent control (DMSO) for 3 h. Then the cells were
treated with TRAIL (20 ng/mL) for 24 h. The whole cell lysates were
collected using PhosphoSafe Buffer (EMD Chemicals). Protein
concentrations were measured with the BCA Protein Assay kit (Thermo
Fisher Scientific). Lysates containing equal amounts of protein
were separated by NuPAGE 4-12% Bis-Tris protein gels (Thermo Fisher
Scientific), transferred to polyvinylidene difluoride membranes,
probed with primary and secondary antibodies. The membranes were
visualized using Li-Cor imaging system. Anti-PARP and anti-actin
antibodies were from Cell Signaling. Secondary anti-mouse and
anti-rabbit antibodies were from Invitrogen.
INCORPORATION BY REFERENCE
[0501] The contents of all references (including literature
references, issued patents, published patent applications, and
co-pending patent applications) cited throughout this application
are hereby expressly incorporated herein in their entireties by
reference.
EQUIVALENTS
[0502] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents of the specific embodiments of the invention described
herein. Such equivalents are intended with be encompassed by the
following claims.
* * * * *